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内山圭司

徳島大学

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リサーチマップ・
Jグローバル
KAKEN
注目研究

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2024年4月25日更新

職名: 特任准教授
電話: 088-633-7439
電子メール: ku200@tokushima-u.ac.jp
学歴: 1992/3: 大阪大学工学部醗酵科卒業
1994/3: 大阪大学大学院工学研究科醗酵工学専攻修士課程修了
1997/3: 大阪大学大学院工学研究科醗酵工学専攻博士課程修了
学位: 博士(工学) (大阪大学) (1997年3月)
職歴・経歴: 1997/4: 大阪大学大学院工学研究科応用生物工学専攻助手
2000/3: University of Cambridge, Cambridge Institute for Medical Research, Visiting Researcher
2003/10: University of Cambridge, Cambridge Institute for Medical Research, Senior Research Associate
2004/4: 三菱化学生命科学研究所研究部門細胞構造・病態グループ主任研究員
2007/4: 徳島大学疾患酵素学研究センターシグナル伝達と糖尿病研究部門准教授
2011/4: 徳島大学疾患酵素学研究センター神経変性疾患研究部門准教授
2016/4: 徳島大学先端酵素学研究所神経変性病態学分野准教授
2020/4: 徳島県立総合看護学校第二看護学科非常勤講師(生化学)

専門分野・研究分野: 分子細胞生物学 (Molecular and Cellular Biology)
神経化学 (Neurochemistry)
分子細胞生物学 (Molecular and Cellular Biology)

研究者総覧で最新データを確認する

2024年4月25日更新

専門分野・研究分野: 分子細胞生物学 (Molecular and Cellular Biology)
神経化学 (Neurochemistry)
分子細胞生物学 (Molecular and Cellular Biology)
担当経験のある授業科目: 研究者総覧に該当データはありませんでした。
指導経験: 研究者総覧に該当データはありませんでした。

研究者総覧で最新データを確認する

2024年4月25日更新

専門分野・研究分野: 分子細胞生物学 (Molecular and Cellular Biology)
神経化学 (Neurochemistry)
分子細胞生物学 (Molecular and Cellular Biology)

研究テーマ: プリオン蛋白質による細胞死誘導機構
異常プリオン蛋白質の複製機構
高生産株の構築を目指した抗体生産CHO細胞の分子細胞生物学的研究

著書

S Sakaguchi and Keiji Uchiyama :
Prion Propagation, Its Neurotoxicity, and Protein Trafficking,
Caister Academic Press, Apr. 2019. Keiji Uchiyama and S Sakaguchi :
A Molecular Mechanism for Abnormal Prion Protein Accumulation,
IntechOpen, Nov. 2018. Keiji Uchiyama and Suehiro Sakaguchi :
Immunological strategies for the prevention and treatment of prion diseases.,
Caister Academic Press, Norfolk, UK, 2013.

(キーワード): プリオン

Keiji Uchiyama, T Suzuki, H Tatsumi, H Kanetake and S Shioya :
Amplified 16S Ribosomal DNA Restriction Analysis of Microbial Community Structure During Fast Degradation of a Biopolymer, PHA, by Composting.,
Springer-Verlag, Berlin, 2002. 清水浩, 内山圭司, 永久圭介, 塩谷捨明 :
生物工学実験書第6章環境化学実験, 6.2 河川の水質調査とモデル解析,
日本生物工学会, 2002年.

論文

Keiji Uchiyama, Hideyuki Hara, Junji Chida, Agriani Dini Pasiana, Morikazu Imamura, Tsuyoshi Mori, Hanae Takatsuki, Ryuichiro Atarashi and Suehiro Sakaguchi :
Ethanolamine Is a New Anti-Prion Compound,
International Journal of Molecular Sciences, Vol.22, No.21, 11742, 2021.

(要約): Prion diseases are a group of fatal neurodegenerative disorders caused by accumulation of proteinaceous infectious particles, or prions, which mainly consist of the abnormally folded, amyloidogenic prion protein, designated PrP. PrP is produced through conformational conversion of the cellular isoform of prion protein, PrP, in the brain. To date, no effective therapies for prion diseases have been developed. In this study, we incidentally noticed that mouse neuroblastoma N2a cells persistently infected with 22L scrapie prions, termed N2aC24L1-3 cells, reduced PrP levels when cultured in advanced Dulbecco's modified eagle medium (DMEM) but not in classic DMEM. PrP levels remained unchanged in prion-uninfected parent N2aC24 cells cultured in advanced DMEM. These results suggest that advanced DMEM may contain an anti-prion compound(s). We then successfully identified ethanolamine in advanced DMEM has an anti-prion activity. Ethanolamine reduced PrP levels in N2aC24L1-3 cells, but not PrP levels in N2aC24 cells. Also, oral administration of ethanolamine through drinking water delayed prion disease in mice intracerebrally inoculated with RML scrapie prions. These results suggest that ethanolamine could be a new anti-prion compound.
(キーワード): Animals / Brain / Cell Line, Tumor / Ethanolamine / Mice / Mice, Inbred ICR / PrPSc Proteins / Prion Diseases
(徳島大学機関リポジトリ): ● Metadata: 117551
(出版サイトへのリンク): ● Publication site (DOI): 10.3390/ijms222111742
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 34769172; ● Summary page in Scopus @ Elsevier: 2-s2.0-85118175726

(徳島大学機関リポジトリ: 117551, DOI: 10.3390/ijms222111742, PubMed: 34769172, Elsevier: Scopus) Hideyuki Hara, Junji Chida, Keiji Uchiyama, Dini Agriani Pasiana, Etsuhisa Takahashi, Hiroshi Kido and Suehiro Sakaguchi :
Neurotropic influenza A virus infection causes prion protein misfolding into infectious prions in neuroblastoma cells.,
Scientific Reports, Vol.11, No.1, 10109, 2021.

(徳島大学機関リポジトリ): ● Metadata: 117718
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/s41598-021-89586-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 33980968; ● Search Scopus @ Elsevier (PMID): 33980968; ● Search Scopus @ Elsevier (DOI): 10.1038/s41598-021-89586-6

(徳島大学機関リポジトリ: 117718, DOI: 10.1038/s41598-021-89586-6, PubMed: 33980968) Hideyuki Hara, Junji Chida, Agriani Dini Pasiana, Keiji Uchiyama, Yutaka Kikuchi, Tomoko Naito, Yuichi Takahashi, Junji Yamamura, Hisashi Kuromatsu and Suehiro Sakaguchi :
Vaporized Hydrogen Peroxide and Ozone Gas Synergistically Reduce Prion Infectivity on Stainless Steel Wire.,
International Journal of Molecular Sciences, Vol.22, No.6, 3268, 2021.

(要約): Prions are infectious agents causing prion diseases, which include Creutzfeldt-Jakob disease (CJD) in humans. Several cases have been reported to be transmitted through medical instruments that were used for preclinical CJD patients, raising public health concerns on iatrogenic transmissions of the disease. Since preclinical CJD patients are currently difficult to identify, medical instruments need to be adequately sterilized so as not to transmit the disease. In this study, we investigated the sterilizing activity of two oxidizing agents, ozone gas and vaporized hydrogen peroxide, against prions fixed on stainless steel wires using a mouse bioassay. Mice intracerebrally implanted with prion-contaminated stainless steel wires treated with ozone gas or vaporized hydrogen peroxide developed prion disease later than those implanted with control prion-contaminated stainless steel wires, indicating that ozone gas and vaporized hydrogen peroxide could reduce prion infectivity on wires. Incubation times were further elongated in mice implanted with prion-contaminated stainless steel wires treated with ozone gas-mixed vaporized hydrogen peroxide, indicating that ozone gas mixed with vaporized hydrogen peroxide reduces prions on these wires more potently than ozone gas or vaporized hydrogen peroxide. These results suggest that ozone gas mixed with vaporized hydrogen peroxide might be more useful for prion sterilization than ozone gas or vaporized hydrogen peroxide alone.
(キーワード): Animals / Brain / Disease Models, Animal / Dose-Response Relationship, Drug / Hydrogen Peroxide / Mice / Ozone / PrPC Proteins / Prion Diseases / Prions / Stainless Steel
(徳島大学機関リポジトリ): ● Metadata: 116339
(出版サイトへのリンク): ● Publication site (DOI): 10.3390/ijms22063268
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 33806892; ● Summary page in Scopus @ Elsevier: 2-s2.0-85102845949

(徳島大学機関リポジトリ: 116339, DOI: 10.3390/ijms22063268, PubMed: 33806892, Elsevier: Scopus) Keiji Uchiyama, Miyata Hironori, Yamaguchi Yoshitaka, Imamura Morikazu, Okazaki Mariya, Pasiana Dini Agriani, Junji Chida, Hideyuki Hara, Atarashi Ryuichiro, Watanabe Hitomi, Kondoh Gen and Suehiro Sakaguchi :
Strain-Dependent Prion Infection in Mice Expressing Prion Protein with Deletion of Central Residues 91-106.,
International Journal of Molecular Sciences, Vol.21, No.19, 7260, 2020.

(要約): Conformational conversion of the cellular prion protein, PrP, into the abnormally folded isoform, PrP, is a key pathogenic event in prion diseases. However, the exact conversion mechanism remains largely unknown. Transgenic mice expressing PrP with a deletion of the central residues 91-106 were generated in the absence of endogenous PrP, designated Tg(PrP∆91-106)/ mice and intracerebrally inoculated with various prions. Tg(PrP∆91-106)/ mice were resistant to RML, 22L and FK-1 prions, neither producing PrP∆91-106 or prions in the brain nor developing disease after inoculation. However, they remained marginally susceptible to bovine spongiform encephalopathy (BSE) prions, developing disease after elongated incubation times and accumulating PrP∆91-106 and prions in the brain after inoculation with BSE prions. Recombinant PrP∆91-104 converted into PrP∆91-104 after incubation with BSE-PrP-prions but not with RML- and 22L-PrP-prions, in a protein misfolding cyclic amplification assay. However, digitonin and heparin stimulated the conversion of PrP∆91-104 into PrP∆91-104 even after incubation with RML- and 22L-PrP-prions. These results suggest that residues 91-106 or 91-104 of PrP are crucially involved in prion pathogenesis in a strain-dependent manner and may play a similar role to digitonin and heparin in the conversion of PrP into PrP.
(徳島大学機関リポジトリ): ● Metadata: 115634
(出版サイトへのリンク): ● Publication site (DOI): 10.3390/ijms21197260
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 33019549; ● Summary page in Scopus @ Elsevier: 2-s2.0-85091919621

(徳島大学機関リポジトリ: 115634, DOI: 10.3390/ijms21197260, PubMed: 33019549, Elsevier: Scopus) Junji Chida, Hideyuki Hara, Keiji Uchiyama, Etsuhisa Takahashi, Hironori Miyata, Hidetaka Kosako, Yukiko Tomioka, Toshihiro Ito, Hiroyuki Horiuchi, Haruo Matsuda, Hiroshi Kido and Suehiro Sakaguchi :
Prion protein signaling induces M2 macrophage polarization and protects from lethal influenza infection in mice.,
PLoS Pathogens, Vol.16, No.8, e1008823, 2020.

(要約): The cellular prion protein, PrPC, is a glycosylphosphatidylinositol anchored-membrane glycoprotein expressed most abundantly in neuronal and to a lesser extent in non-neuronal cells. Its conformational conversion into the amyloidogenic isoform in neurons is a key pathogenic event in prion diseases, including Creutzfeldt-Jakob disease in humans and scrapie and bovine spongiform encephalopathy in animals. However, the normal functions of PrPC remain largely unknown, particularly in non-neuronal cells. Here we show that stimulation of PrPC with anti-PrP monoclonal antibodies (mAbs) protected mice from lethal infection with influenza A viruses (IAVs), with abundant accumulation of anti-inflammatory M2 macrophages with activated Src family kinases (SFKs) in infected lungs. A SFK inhibitor dasatinib inhibited M2 macrophage accumulation in IAV-infected lungs after treatment with anti-PrP mAbs and abolished the anti-PrP mAb-induced protective activity against lethal influenza infection in mice. We also show that stimulation of PrPC with anti-PrP mAbs induced M2 polarization in peritoneal macrophages through SFK activation in vitro and in vivo. These results indicate that PrPC could activate SFK in macrophages and induce macrophage polarization to an anti-inflammatory M2 phenotype after stimulation with anti-PrP mAbs, thereby eliciting protective activity against lethal infection with IAVs in mice after treatment with anti-PrP mAbs. These results also highlight PrPC as a novel therapeutic target for IAV infection.
(徳島大学機関リポジトリ): ● Metadata: 115613
(出版サイトへのリンク): ● Publication site (DOI): 10.1371/journal.ppat.1008823
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 32845931; ● Search Scopus @ Elsevier (PMID): 32845931; ● Search Scopus @ Elsevier (DOI): 10.1371/journal.ppat.1008823

(徳島大学機関リポジトリ: 115613, DOI: 10.1371/journal.ppat.1008823, PubMed: 32845931) Nandita Rani Das, Hironori Miyata, Hideyuki Hara, Junji Chida, Keiji Uchiyama, Kentaro Masujin, Hitomi Watanabe, Gen Kondoh and Suehiro Sakaguchi :
The N-Terminal Polybasic Region of Prion Protein Is Crucial in Prion Pathogenesis Independently of the Octapeptide Repeat Region,
Molecular Neurobiology, Vol.57, 1203-1216, 2019.

(要約): Conformational conversion of the cellular isoform of prion protein, designated PrP, into the abnormally folded, amyloidogenic isoform, PrP, is an essential pathogenic event in prion diseases. However, the exact conversion mechanism remains largely unknown. Lines of evidence indicate that the N-terminal domain, which includes the N-terminal, positively charged polybasic region and the octapeptide repeat (OR) region, is important for PrP to convert into PrP after infection with prions. To further gain insights into the role of the polybasic region and the OR region in prion pathogenesis, we generated two different transgenic mice, designated Tg(PrP3K3A)/Prnp and Tg(PrP3K3A∆OR)/Prnp mice, which express PrP with lysine residues at codons 23, 24, and 27 in the polybasic region mutated with or without a deletion of the OR region on the Prnp background, respectively, and intracerebrally inoculated them with RML and 22L prions. We show that Tg(PrP3K3A)/Prnp mice were highly resistant to the prions, indicating that lysine residues at 23, 24, and 27 could be important for the polybasic region to support prion infection. Tg(PrP3K3A∆OR)/Prnp mice also had reduced susceptibility to RML and 22L prions equivalent to Tg(PrP3K3A)/Prnp mice. The pre-OR region, including the polybasic region, of PrP3K3A∆OR, but not PrP3K3A, was unusually converted to a protease-resistant structure during conversion to PrP3K3A∆OR. These results suggest that, while the OR region could affect the conformation of the polybasic region during conversion of PrP into PrP, the polybasic region could play a crucial role in prion pathogenesis independently of the OR region.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s12035-019-01804-5
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 31707632; ● Summary page in Scopus @ Elsevier: 2-s2.0-85074844020

(DOI: 10.1007/s12035-019-01804-5, PubMed: 31707632, Elsevier: Scopus) Kohei Kaneyoshi, Noriko Yamano-Adachi, Yuichi Koga, Keiji Uchiyama and Takeshi Omasa :
Analysis of the immunoglobulin G (IgG) secretion efficiency in recombinant Chinese hamster ovary (CHO) cells by using Citrine-fusion IgG.,
Cytotechnology, Vol.71, No.1, 193-207, 2019.

(要約): Biopharmaceuticals represented by immunoglobulin G (IgG) are produced by the cultivation of recombinant animal cells, especially Chinese hamster ovary (CHO) cells. It is thought that the intracellular secretion process of IgG is a bottleneck in the production of biopharmaceuticals. Many studies on the regulation of endogenous secretory protein expression levels have shown improved productivity. However, these strategies have not universally improved the productivity of various proteins. A more rational and efficient establishment of high producer cells is required based on an understanding of the secretory processes in IgG producing CHO cells. In this study, a CHO cell line producing humanized IgG1, which was genetically fused with fluorescent proteins, was established to directly analyze intracellular secretion. The relationship between the amount of intracellular and secreted IgG was analyzed at the single cell level by an automated single-cell analysis and isolation system equipped with dual color fluorescent filters. The amounts of intracellular and secreted IgG showed a weak positive correlation. The amount of secreted IgG analyzed by the system showed a weak negative linear correlation with the specific growth of isolated clones. An immunofluorescent microscopy study showed that the established clones could be used to analyze the intracellular secretion bottleneck. This is the first study to report the use of fluorescent protein fusion IgG as a tool to analyze the secretion of recombinant CHO cells.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s10616-018-0276-7
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 30610509; ● Summary page in Scopus @ Elsevier: 2-s2.0-85059533371

(DOI: 10.1007/s10616-018-0276-7, PubMed: 30610509, Elsevier: Scopus) Kohei Kaneyoshi, Kouki Kuroda, Keiji Uchiyama, Masayoshi Onitsuka, Noriko Yamano-Adachi, Yuichi Koga and Takeshi Omasa :
Secretion analysis of intracellular difficult-to-express immunoglobulin G (IgG) in Chinese hamster ovary (CHO) cells,
Cytotechnology, Vol.71, No.1, 305-316, 2019.

(要約): The Chinese hamster ovary (CHO) cell line is the most widely used host cell for therapeutic antibody production. Although its productivity has been improved by various strategies to satisfy the growing global demand, some difficult-to-express (DTE) antibodies remain at low secretion levels. To improve the production of various therapeutic antibodies, it is necessary to determine possible rate-limiting steps in DTE antibody secretion in comparison with other high IgG producers. Here, we analyzed the protein secretion process in CHO cells producing the DTE immunoglobulin G (IgG) infliximab. The results from chase assays using a translation inhibitor revealed that infliximab secretion could be nearly completed within 2 h, at which time the cells still retained about 40% of heavy chains and 65% of light chains. Using fluorescent microscopy, we observed that these IgG chains remained in the endoplasmic reticulum and Golgi apparatus. The cells inefficiently form fully assembled heterodimer IgG by making LC aggregates, which may be the most serious bottleneck in the production of DTE infliximab compared with other IgG high producers. Our study could contribute to establish the common strategy for constructing DTE high-producer cells on the basis of rate-limiting step analysis.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s10616-018-0286-5
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 30637508; ● Summary page in Scopus @ Elsevier: 2-s2.0-85060048909

(DOI: 10.1007/s10616-018-0286-5, PubMed: 30637508, Elsevier: Scopus) Kohei Kaneyoshi, Keiji Uchiyama, Masayoshi Onitsuka, Noriko Yamano, Yuichi Koga and Takeshi Omasa :
Analysis of intracellular IgG secretion in Chinese hamster ovary cells to improve IgG production.,
Journal of Bioscience and Bioengineering, Vol.127, No.1, 107-113, 2019.

(要約): The production of biopharmaceutical immunoglobulin G (IgG) using cultured mammalian cells, especially Chinese hamster ovary (CHO) cells is well established and has been markedly improved through the modification of cells and cell culture engineering technologies. The establishment of high-production cell lines remains a challenge. The intracellular secretion of IgG has been investigated to identify and solve the rate-limiting steps in antibody production. However, strategies that regulate the expression of proteins that are related to antibody secretory pathway have not consistently improved their production. In this study, key features and limitations of the antibody secretion process in recombinant CHO cells were analyzed to develop more efficient approaches for establishing high-production cells. By chase assay with protein translation inhibitors, IgG secretion reached a plateau when at least 20% of IgG remained in the cells. The secretion kinetics and retention ratio of IgG varied between IgG subclasses (two types of IgG1 and an IgG3 subclass). Immunofluorescent microscopy and size exclusion chromatography showed that the remaining intracellular IgG localized mainly within the endoplasmic reticulum (ER) and less with the cis-Golgi network, despite the formation of fully assembled IgG. These results show that remaining intracellular IgG is a target for enhancing antibody secretion, even in high-production CHO cells.
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2018.06.018
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 30017708; ● Search Scopus @ Elsevier (PMID): 30017708; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2018.06.018

(DOI: 10.1016/j.jbiosc.2018.06.018, PubMed: 30017708) Junji Chida, Hideyuki Hara, Masashi Yano, Keiji Uchiyama, Rani Nandita Das, Etsuhisa Takahashi, Hironori Miyata, Yukiko Tomioka, Toshihiro Ito, Hiroshi Kido and Suehiro Sakaguchi :
Prion protein protects mice from lethal infection with influenza A viruses.,
PLoS Pathogens, Vol.14, No.5, e1007049, 2018.

(要約): The cellular prion protein, designated PrPC, is a membrane glycoprotein expressed abundantly in brains and to a lesser extent in other tissues. Conformational conversion of PrPC into the amyloidogenic isoform is a key pathogenic event in prion diseases. However, the physiological functions of PrPC remain largely unknown, particularly in non-neuronal tissues. Here, we show that PrPC is expressed in lung epithelial cells, including alveolar type 1 and 2 cells and bronchiolar Clara cells. Compared with wild-type (WT) mice, PrPC-null mice (Prnp0/0) were highly susceptible to influenza A viruses (IAVs), with higher mortality. Infected Prnp0/0 lungs were severely injured, with higher inflammation and higher apoptosis of epithelial cells, and contained higher reactive oxygen species (ROS) than control WT lungs. Treatment with a ROS scavenger or an inhibitor of xanthine oxidase (XO), a major ROS-generating enzyme in IAV-infected lungs, rescued Prnp0/0 mice from the lethal infection with IAV. Moreover, Prnp0/0 mice transgenic for PrP with a deletion of the Cu-binding octapeptide repeat (OR) region, Tg(PrPOR)/Prnp0/0 mice, were also highly susceptible to IAV infection. These results indicate that PrPC has a protective role against lethal infection with IAVs through the Cu-binding OR region by reducing ROS in infected lungs. Cu content and the activity of anti-oxidant enzyme Cu/Zn-dependent superoxide dismutase, SOD1, were lower in Prnp0/0 and Tg(PrPOR)/Prnp0/0 lungs than in WT lungs. It is thus conceivable that PrPC functions to maintain Cu content and regulate SOD1 through the OR region in lungs, thereby reducing ROS in IAV-infected lungs and eventually protecting them from lethal infection with IAVs. Our current results highlight the role of PrPC in protection against IAV infection, and suggest that PrPC might be a novel target molecule for anti-influenza therapeutics.
(キーワード): Animals / Brain / Copper / Disease Susceptibility / Influenza A virus / Lung / Mice / Mice, Inbred C57BL / Mice, Knockout / Neurons / PrPC Proteins / Prion Diseases / Prion Proteins / Prions / Reactive Oxygen Species / Superoxide Dismutase
(徳島大学機関リポジトリ): ● Metadata: 112206
(出版サイトへのリンク): ● Publication site (DOI): 10.1371/journal.ppat.1007049
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 29723291; ● Search Scopus @ Elsevier (PMID): 29723291; ● Search Scopus @ Elsevier (DOI): 10.1371/journal.ppat.1007049

(徳島大学機関リポジトリ: 112206, DOI: 10.1371/journal.ppat.1007049, PubMed: 29723291) Luise Linsenmeier, Behnam Mohammadi, Sebastian Wetzel, Berta Puig, Walker S. Jackson, Alexander Hartmann, Keiji Uchiyama, Suehiro Sakaguchi, Kristina Endres, Jörg Tatzelt, Paul Saftig, Markus Glatzel and Hermann C. Altmeppen :
Structural and mechanistic aspects influencing the ADAM10-mediated shedding of the prion protein.,
Molecular Neurodegeneration, Vol.13, No.1, 18, 2018.

(要約): Proteolytic processing of the prion protein (PrP<superscript>C</superscript>) by endogenous proteases generates bioactive membrane-bound and soluble fragments which may help to explain the pleiotropic roles of this protein in the nervous system and in brain diseases. Shedding of almost full-length PrP<superscript>C</superscript> into the extracellular space by the metalloprotease ADAM10 is of peculiar relevance since soluble PrP stimulates axonal outgrowth and is protective in neurodegenerative conditions such as Alzheimer's and prion disease. However, molecular determinates and mechanisms regulating the shedding of PrP are entirely unknown. We produced an antibody recognizing the neo-epitope of shed PrP generated by ADAM10 in biological samples and used it to study structural and mechanistic aspects affecting the shedding. For this, we investigated genetically modified cellular and murine models by biochemical and morphological approaches. We show that the novel antibody specifically detects shed PrP in cell culture supernatants and murine brain. We demonstrate that ADAM10 is the exclusive sheddase of PrP<superscript>C</superscript> in the nervous system and reveal that the glycosylation state and type of membrane-anchorage of PrP<superscript>C</superscript> severely affect its shedding. Furthermore, we provide evidence that PrP shedding can be modulated by pharmacological inhibition and stimulation and present data suggesting that shedding is a relevant part of a compensatory network ensuring PrP<superscript>C</superscript> homeostasis of the cell. With the new antibody, our study introduces a new tool to reliably investigate PrP-shedding. In addition, this study provides novel and important insight into the regulation of this cleavage event, which is likely to be relevant for diagnostic and therapeutic approaches even beyond neurodegeneration.
(キーワード): ADAM10 Protein / Animals / 脳 (brain) / Mice / Neurons / Prion Proteins
(徳島大学機関リポジトリ): ● Metadata: 114545
(出版サイトへのリンク): ● Publication site (DOI): 10.1186/s13024-018-0248-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 29625583; ● Summary page in Scopus @ Elsevier: 2-s2.0-85044987648

(徳島大学機関リポジトリ: 114545, DOI: 10.1186/s13024-018-0248-6, PubMed: 29625583, Elsevier: Scopus) Hideyuki Hara, Miyata Hironori, Das Rani Nandita, Junji Chida, Yoshimochi Tatenobu, Keiji Uchiyama, Watanabe Hitomi, Kondoh Gen, Yokoyama Takashi and Suehiro Sakaguchi :
Prion Protein Devoid of the Octapeptide Repeat Region Delays BSE Pathogenesis in Mice.,
Journal of Virology, Vol.92, No.1, pii:e01368-17, 2018.

(要約): Conformational conversion of the cellular isoform of prion protein, PrP, into the abnormally folded, amyloidogenic isoform, PrP, is a key pathogenic event in prion diseases, including Creutzfeldt-Jakob disease in humans and scrapie and bovine spongiform encephalopathy (BSE) in animals. We previously reported that the octapeptide repeat (OR) region could be dispensable for converting PrP into PrP after infection with RML prions. We demonstrated that mice transgenically expressing mouse PrP with deletion of the OR region on the PrP knockout background, designated Tg(PrPΔOR)/ mice, did not show reduced susceptibility to RML scrapie prions, with abundant accumulation of PrPΔOR in their brains. We show here that Tg(PrPΔOR)/ mice were highly resistant to BSE prions, developing the disease with markedly elongated incubation times after infection with BSE prions. The conversion of PrPΔOR into PrPΔOR was markedly delayed in their brains. These results suggest that the OR region may have a crucial role in the conversion of PrP into PrP after infection with BSE prions. However, Tg(PrPΔOR)/ mice remained susceptible to RML and 22L scrapie prions, developing the disease without elongated incubation times after infection with RML and 22L prions. PrPΔOR accumulated only slightly less in the brains of RML- or 22L-infected Tg(PrPΔOR)/ mice than PrP in control wild-type mice. Taken together, these results indicate that the OR region of PrP could play a differential role in the pathogenesis of BSE prions and RML or 22L scrapie prions. Structure-function relationship studies of PrP conformational conversion into PrP are worthwhile to understand the mechanism of the conversion of PrP into PrP We show here that, by inoculating Tg(PrPΔOR)/ mice with the three different strains of RML, 22L, and BSE prions, the OR region could play a differential role in the conversion of PrP into PrP after infection with RML or 22L scrapie prions and BSE prions. PrPΔOR was efficiently converted into PrPΔOR after infection with RML and 22L prions. However, the conversion of PrPΔOR into PrPΔOR was markedly delayed after infection with BSE prions. Further investigation into the role of the OR region in the conversion of PrP into PrP after infection with BSE prions might be helpful for understanding the pathogenesis of BSE prions.
(キーワード): Animals / Brain / Cattle / Disease Susceptibility / Encephalopathy, Bovine Spongiform / Humans / Mice / Mice, Transgenic / Oligopeptides / PrPC Proteins / Prion Diseases / Prions / Sequence Deletion
(徳島大学機関リポジトリ): ● Metadata: 111465
(出版サイトへのリンク): ● Publication site (DOI): 10.1128/JVI.01368-17
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 29046443; ● CiNii @ 国立情報学研究所 (CRID): 1050845762396495104; ● Search Scopus @ Elsevier (PMID): 29046443; ● Search Scopus @ Elsevier (DOI): 10.1128/JVI.01368-17

(徳島大学機関リポジトリ: 111465, DOI: 10.1128/JVI.01368-17, PubMed: 29046443, CiNii: 1050845762396495104) Suehiro Sakaguchi and Keiji Uchiyama :
Novel Amplification Mechanism of Prions through Disrupting Sortilin-Mediated Trafficking.,
Prion, Vol.11, No.6, 398-404, 2017.

(要約): Conformational conversion of the cellular prion protein, PrP, into the abnormally folded isoform of prion protein, PrP, which leads to marked accumulation of PrP in brains, is a key pathogenic event in prion diseases, a group of fatal neurodegenerative disorders caused by prions. However, the exact mechanism of PrP accumulation in prion-infected neurons remains unknown. We recently reported a novel cellular mechanism to support PrP accumulation in prion-infected neurons, in which PrP itself promotes its accumulation by evading the cellular inhibitory mechanism, which is newly identified in our recent study. We showed that the VPS10P sorting receptor sortilin negatively regulates PrP accumulation in prion-infected neurons, by interacting with PrP and PrP and trafficking them to lysosomes for degradation. However, PrP stimulated lysosomal degradation of sortilin, disrupting the sortilin-mediated degradation of PrP and PrP and eventually evoking further accumulation of PrP in prion-infected neurons. These findings suggest a positive feedback amplification mechanism for PrP accumulation in prion-infected neurons.
(徳島大学機関リポジトリ): ● Metadata: 112200
(出版サイトへのリンク): ● Publication site (DOI): 10.1080/19336896.2017.1391435
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 29099278; ● Summary page in Scopus @ Elsevier: 2-s2.0-85034055921

(徳島大学機関リポジトリ: 112200, DOI: 10.1080/19336896.2017.1391435, PubMed: 29099278, Elsevier: Scopus) Keiji Uchiyama, Mitsuru Tomita, Masashi Yano, Junji Chida, Hideyuki Hara, Nandita Rani Das, Anders Nykjaer and Suehiro Sakaguchi :
Prions amplify through degradation of the VPS10P sorting receptor sortilin.,
PLoS Pathogens, Vol.13, No.6, e1006470, 2017.

(要約): Prion diseases are a group of fatal neurodegenerative disorders caused by prions, which consist mainly of the abnormally folded isoform of prion protein, PrPSc. A pivotal pathogenic event in prion disease is progressive accumulation of prions, or PrPSc, in brains through constitutive conformational conversion of the cellular prion protein, PrPC, into PrPSc. However, the cellular mechanism by which PrPSc is progressively accumulated in prion-infected neurons remains unknown. Here, we show that PrPSc is progressively accumulated in prion-infected cells through degradation of the VPS10P sorting receptor sortilin. We first show that sortilin interacts with PrPC and PrPSc and sorts them to lysosomes for degradation. Consistently, sortilin-knockdown increased PrPSc accumulation in prion-infected cells. In contrast, overexpression of sortilin reduced PrPSc accumulation in prion-infected cells. These results indicate that sortilin negatively regulates PrPSc accumulation in prion-infected cells. The negative role of sortilin in PrPSc accumulation was further confirmed in sortilin-knockout mice infected with prions. The infected mice had accelerated prion disease with early accumulation of PrPSc in their brains. Interestingly, sortilin was reduced in prion-infected cells and mouse brains. Treatment of prion-infected cells with lysosomal inhibitors, but not proteasomal inhibitors, increased the levels of sortilin. Moreover, sortilin was reduced following PrPSc becoming detectable in cells after infection with prions. These results indicate that PrPSc accumulation stimulates sortilin degradation in lysosomes. Taken together, these results show that PrPSc accumulation of itself could impair the sortilin-mediated sorting of PrPC and PrPSc to lysosomes for degradation by stimulating lysosomal degradation of sortilin, eventually leading to progressive accumulation of PrPSc in prion-infected cells.
(キーワード): prion / Sortilin / VPS10P
(徳島大学機関リポジトリ): ● Metadata: 110433
(出版サイトへのリンク): ● Publication site (DOI): 10.1371/journal.ppat.1006470
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 28665987; ● Search Scopus @ Elsevier (PMID): 28665987; ● Search Scopus @ Elsevier (DOI): 10.1371/journal.ppat.1006470

(徳島大学機関リポジトリ: 110433, DOI: 10.1371/journal.ppat.1006470, PubMed: 28665987) Das Rani Nandita, Miyata Hironori, Hideyuki Hara, Keiji Uchiyama, Junji Chida, Masashi Yano, Watanabe Hitomi, Kondoh Gen and Suehiro Sakaguchi :
Effects of prion protein devoid of the N-terminal residues 25-50 on prion pathogenesis in mice.,
Archives of Virology, Vol.162, No.7, 1867-1876, 2017.

(要約): The N-terminal polybasic region of the normal prion protein, PrP(C), which encompasses residues 23-31, is important for prion pathogenesis by affecting conversion of PrP(C) into the pathogenic isoform, PrP(Sc). We previously reported transgenic mice expressing PrP with residues 25-50 deleted in the PrP-null background, designated as Tg(PrPpreOR)/Prnp (0/0) mice. Here, we produced two new lines of Tg(PrPpreOR)/Prnp (0/0) mice, each expressing the mutant protein, PrPpreOR, 1.1 and 1.6 times more than PrP(C) in wild-type mice, and subsequently intracerebrally inoculated RML and 22L prions into them. The lower expresser showed slightly reduced susceptibility to RML prions but not to 22L prions. The higher expresser exhibited enhanced susceptibility to both prions. No prion transmission barrier was created in Tg(PrPpreOR)/Prnp (0/0) mice against full-length PrP(Sc). PrP(Sc)preOR accumulated in the brains of infected Tg(PrPpreOR)/Prnp (0/0) mice less than PrP(Sc) in control wild-type mice, although lower in RML-infected Tg(PrPpreOR)/Prnp (0/0) mice than in 22L-infected mice. Prion infectivity in infected Tg(PrPpreOR)/Prnp (0/0) mice was also lower than that in wild-type mice. These results indicate that deletion of residues 25-50 only slightly affects prion susceptibility, the conversion of PrP(C) into PrP(Sc), and prion infectivity in a strain-specific way. PrPpreOR retains residues 23-24 and lacks residues 25-31 in the polybasic region. It is thus conceivable that residues 23-24 rather than 25-31 are important for the polybasic region to support prion pathogenesis. However, other investigators have reported that residues 27-31 not 23-24 are important to support prion pathogenesis. Taken together, the polybasic region might support prion pathogenesis through multiple sites including residues 23-24 and 27-31.
(キーワード): Amino Acid Sequence / Animals / Disease Susceptibility / Mice / Mice, Inbred C57BL / Mice, Transgenic / Prion Diseases / Prion Proteins / Repetitive Sequences, Amino Acid / Sequence Deletion
(徳島大学機関リポジトリ): ● Metadata: 110119
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00705-017-3295-3
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 28255815; ● Summary page in Scopus @ Elsevier: 2-s2.0-85014121673

(徳島大学機関リポジトリ: 110119, DOI: 10.1007/s00705-017-3295-3, PubMed: 28255815, Elsevier: Scopus) Keiji Uchiyama, Miyata Hironori, Masashi Yano, Yoshitaka Yamaguti, Imamura Morikazu, Muramatsu Naomi, Das Rani Nandita, Junji Chida, Hideyuki Hara and Suehiro Sakaguchi :
Mouse-Hamster Chimeric Prion Protein (PrP) Devoid of N-terminal Residues 23-88 Restores Susceptibility to 22L Prions, But Not to RML Prions in PrP-Knockout Mice.,
PLoS ONE, Vol.9, No.10, e109737, 2014.

(要約): Prion infection induces conformational conversion of the normal prion protein PrPC, into the pathogenic isoform PrPSc, in prion diseases. It has been shown that PrP-knockout (Prnp0/0) mice transgenically reconstituted with a mouse-hamster chimeric PrP lacking N-terminal residues 23-88, or Tg(MHM223-88)/Prnp 0/0 mice, neither developed the disease nor accumulated MHM2Sc23-88 in their brains after inoculation with RML prions. In contrast, RML-inoculated Tg(MHM223-88)/Prnp 0/+ mice developed the disease with abundant accumulation of MHM2Sc23-88 in their brains. These results indicate that MHM223-88 itself might either lose or greatly reduce the converting capacity to MHM2Sc23-88, and that the co-expressing wild-type PrPC can stimulate the conversion of MHM223-88 to MHM2Sc23-88 in trans. In the present study, we confirmed that Tg(MHM223-88)/Prnp 0/0 mice remained resistant to RML prions for up to 730 days after inoculation. However, we found that Tg(MHM223-88)/Prnp 0/0 mice were susceptible to 22L prions, developing the disease with prolonged incubation times and accumulating MHM2Sc23-88 in their brains. We also found accelerated conversion of MHM223-88 into MHM2Sc23-88 in the brains of RML- and 22L-inoculated Tg(MHM223-88)/Prnp 0/+ mice. However, wild-type PrPSc accumulated less in the brains of these inoculated Tg(MHM223-88)/Prnp 0/+ mice, compared with RML- and 22L-inoculated Prnp 0/+ mice. These results show that MHM223-88 itself can convert into MHM2Sc23-88 without the help of the trans-acting PrPC, and that, irrespective of prion strains inoculated, the co-expressing wild-type PrPC stimulates the conversion of MHM223-88 into MHM2Sc23-88, but to the contrary, the co-expressing MHM223-88 disturbs the conversion of wild-type PrPC into PrPSc.
(出版サイトへのリンク): ● Publication site (DOI): 10.1371/journal.pone.0109737
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 25330286; ● Search Scopus @ Elsevier (PMID): 25330286; ● Search Scopus @ Elsevier (DOI): 10.1371/journal.pone.0109737

(DOI: 10.1371/journal.pone.0109737, PubMed: 25330286) Tomoyuki Yuasa, Kikuko Amo, Shuhei Ishikura, Hisao Nagaya, Keiji Uchiyama, Seiichi Hashida and Yousuke Ebina :
Development of in vitro model of insulin receptor cleavage induced by high glucose in HepG2 cells.,
Biochemical and Biophysical Research Communications, Vol.445, No.1, 236-243, 2014.

(要約): Soluble insulin receptor (sIR), the ectodomain of IR, has been detected in human plasma, and its concentration parallels that of blood glucose in patients with diabetes. IR has a pivotal role in glucose homeostasis and diabetes development; therefore, cleavage of IR promoted by hyperglycemia is involved in insulin resistance and glucose toxicity. To elucidate the physiology of sIR, we developed an in vitro model mimicking the changes in sIR levels in plasma from patients with diabetes. Among four human cell lines that expressed IR, spontaneous cleavage of IR occurred only in HepG2 cells. The molecular characteristics of sIR derived from HepG2 cells were similar to those of sIR detected in human plasma. The concentration of sIR in the medium did not differ between basal and high-glucose conditions in the initial 24-h period, but increasing the duration of pre-stimulation (>48 h) led to a significant increase in sIR levels in cells exposed to high glucose. Additionally, glucose-dependent increment of sIR was reversible in this model. These results are consistent with the observation of plasma sIR in patients with diabetes. Using this model, O-linked N-acetylglucosamine modification was determined to be involved in high-glucose-induced IR cleavage. A calcium-dependent protease was shown to cleave IR extracellularly. These findings show that this in vitro model could be useful for determining the molecular mechanism underlying IR cleavage.
(キーワード): soluble insulin receptor / Insulin resistance / shedding / O-GlcNAc / Calcium dependent protease / Diabetes / Hep G2
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.bbrc.2014.01.187
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24508798; ● Summary page in Scopus @ Elsevier: 2-s2.0-84896697427

(DOI: 10.1016/j.bbrc.2014.01.187, PubMed: 24508798, Elsevier: Scopus) Keiji Uchiyama, Miyata Hironori and Suehiro Sakaguchi :
Disturbed vesicular trafficking of membrane proteins in prion disease.,
Prion, Vol.7, No.6, 447-451, 2013.

(要約): The pathogenic mechanism of prion diseases remains unknown. We recently reported that prion infection disturbs post-Golgi trafficking of certain types of membrane proteins to the cell surface, resulting in reduced surface expression of membrane proteins and abrogating the signal from the proteins. The surface expression of the membrane proteins was reduced in the brains of mice inoculated with prions, well before abnormal symptoms became evident. Prions or pathogenic prion proteins were mainly detected in endosomal compartments, being particularly abundant in recycling endosomes. Some newly synthesized membrane proteins are delivered to the surface from the Golgi apparatus through recycling endosomes, and some endocytosed membrane proteins are delivered back to the surface through recycling endosomes. These results suggest that prions might cause neuronal dysfunctions and cell loss by disturbing post-Golgi trafficking of membrane proteins via accumulation in recycling endosomes. Interestingly, it was recently shown that delivery of a calcium channel protein to the cell surface was impaired and its function was abrogated in a mouse model of hereditary prion disease. Taken together, these results suggest that impaired delivery of membrane proteins to the cell surface is a common pathogenic event in acquired and hereditary prion diseases.
(出版サイトへのリンク): ● Publication site (DOI): 10.4161/pri.27381
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24335150; ● Summary page in Scopus @ Elsevier: 2-s2.0-84893871953

(DOI: 10.4161/pri.27381, PubMed: 24335150, Elsevier: Scopus) Keiji Uchiyama, Naomi Muramatsu, Masashi Yano, Takeshi Usui, Hironori Miyata and Suehiro Sakaguchi :
Prions disturb post-Golgi trafficking of membrane proteins.,
Nature Communications, Vol.4, 1846, 2013.

(要約): Conformational conversion of normal cellular prion protein PrP(C) into pathogenic PrP(Sc) is central to the pathogenesis of prion diseases. However, the pathogenic mechanism remains unknown. Here we show that post-Golgi vesicular trafficking is significantly delayed in prion-infected N2a cells. Accordingly, cell surface expression of membrane proteins examined, including PrP(C), insulin receptor involved in neuroprotection, and attractin, whose mutation causes prion disease-like spongiform neurodegeneration, is reduced. Instead, they accumulate in the Golgi apparatus. PrP(Sc) is detected throughout endosomal compartments, being particularly abundant in recycling endosome. We also show reduced surface expression of PrP(C) and insulin receptor in prion-infected mouse brains well before the onset of disease. These results suggest that prion infection might impair post-Golgi trafficking of membrane proteins to the cell surface in neurons via PrP(Sc) accumulated in recycling endosome, and eventually induce neuronal dysfunctions associated with prion diseases.
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/ncomms2873
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23673631; ● Search Scopus @ Elsevier (PMID): 23673631; ● Search Scopus @ Elsevier (DOI): 10.1038/ncomms2873

(DOI: 10.1038/ncomms2873, PubMed: 23673631) Yoshitaka Yamaguti, Hironori Miyata, Keiji Uchiyama, Akira Ootsuyama, Sachiko Inubushi, Tsuyoshi Mori, Naomi Muramatsu, Shigeru Katamine and Suehiro Sakaguchi :
Biological and biochemical characterization of mice expressing prion protein devoid of the octapeptide repeat region after infection with prions.,
PLoS ONE, Vol.7, No.8, e43540, 2012.

(要約): Accumulating lines of evidence indicate that the N-terminal domain of prion protein (PrP) is involved in prion susceptibility in mice. In this study, to investigate the role of the octapeptide repeat (OR) region alone in the N-terminal domain for the susceptibility and pathogenesis of prion disease, we intracerebrally inoculated RML scrapie prions into tg(PrPOR)/Prnp(0/0) mice, which express mouse PrP missing only the OR region on the PrP-null background. Incubation times of these mice were not extended. Protease-resistant PrPOR, or PrP(Sc)OR, was easily detectable but lower in the brains of these mice, compared to that in control wild-type mice. Consistently, prion titers were slightly lower and astrogliosis was milder in their brains. However, in their spinal cords, PrP(Sc)OR and prion titers were abundant and astrogliosis was as strong as in control wild-type mice. These results indicate that the role of the OR region in prion susceptibility and pathogenesis of the disease is limited. We also found that the PrP(Sc)OR, including the pre-OR residues 23-50, was unusually protease-resistant, indicating that deletion of the OR region could cause structural changes to the pre-OR region upon prion infection, leading to formation of a protease-resistant structure for the pre-OR region.
(キーワード): Amino Acid Sequence / Animals / Disease Susceptibility / Endopeptidase K / Forelimb / Gene Expression Regulation / Lysine / Mice / Mice, Inbred C57BL / Mice, Transgenic / Oligopeptides / Paresis / Prions / Repetitive Sequences, Amino Acid / Scrapie / Sequence Deletion
(出版サイトへのリンク): ● Publication site (DOI): 10.1371/journal.pone.0043540
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 22927985; ● Search Scopus @ Elsevier (PMID): 22927985; ● Search Scopus @ Elsevier (DOI): 10.1371/journal.pone.0043540

(DOI: 10.1371/journal.pone.0043540, PubMed: 22927985) G. Totsukawa, Y. Kaneko, Keiji Uchiyama, H. Toh, K. Tamura and H. Kondo :
VCIP135 deubiquitinase and its binding protein, WAC, in p97ATPase-mediated membrane fusion.,
The EMBO Journal, Vol.30, No.17, 3581-3593, 2011.

(要約): Two distinct p97 membrane fusion pathways are required for Golgi biogenesis: the p97/p47 and p97/p37 pathways. VCIP135 is necessary for both pathways, while its deubiquitinating activity is required only for the p97/p47 pathway. We have now identified a novel VCIP135-binding protein, WAC. WAC localizes to the Golgi as well as the nucleus. In Golgi membranes, WAC is involved in a complex containing VCIP135 and p97. WAC directly binds to VCIP135 and increases its deubiquitinating activity. siRNA experiments revealed that WAC is required for Golgi biogenesis. In an in vitro Golgi reformation assay, WAC was necessary only for p97/p47-mediated Golgi reassembly, but not for p97/p37-mediated reassembly. WAC is hence thought to function in p97/p47-mediated Golgi membrane fusion by activating the deubiquitinating function of VCIP135. We also showed that the two p97 pathways function in ER membrane fusion as well. An in vitro ER reformation assay revealed that both pathways required VCIP135 but not its deubiquitinating activity for their ER membrane fusion. This was consistent with the finding that WAC is unnecessary for p97-mediated ER membrane fusion.
(キーワード): Adenosine Triphosphatases / Animals / Carrier Proteins / Endopeptidases / Golgi Apparatus / Liver / Membrane Fusion / Membrane Proteins / Nuclear Proteins / Protein Binding / Rats / Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/emboj.2011.260
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 21811234; ● Search Scopus @ Elsevier (PMID): 21811234; ● Search Scopus @ Elsevier (DOI): 10.1038/emboj.2011.260

(DOI: 10.1038/emboj.2011.260, PubMed: 21811234) Tomoyuki Yuasa, Keiji Uchiyama, OGURA Yuko, Masafumi Kimura, TESHIGAWARA Kiyoshi, Toshio Hosaka, TANAKA Yoshinori, Toshiyuki Obata, SANO Hiroyuki, Kazuhiro Kishi and Yousuke Ebina :
The Rab GTPase-Activating Protein AS160 as a Common Regulator of Insulin- and Gαq-Mediated Intracellular GLUT4 Vesicle Distribution,
Endocrine Journal, Vol.56, No.3, 345-359, 2009.

(要約): Akt substrate of 160kDa (AS160) is a Rab GTPase activating protein (GAP) and was recently identified as a component of the insulin signaling pathway of glucose transporter type 4 (GLUT4) translocation. We and others, previously reported that the activation of Galphaq protein-coupled receptors (GalphaqPCRs) also stimulated GLUT4 translocation and glucose uptake in several cell lines. Here, we report that the activation of GalphaqPCRs also promoted phosphorylation of AS160 by the 5'-AMP activated protein kinase (AMPK). The suppression of AS160 phosphorylation by the siRNA mediated AMPKalpha1 subunit knockdown promoted GLUT4 vesicle retention in intracellular compartments. This suppression did not affect the ratio of non-induced cell surface GLUT4 to Galphaq-induced it. Rat 3Y1 cells lacking AS160 did not show insulin-induced GLUT4 translocation. The cells stably expressing GLUT4 revealed GLUT4 vesicles that were mainly localized in the perinuclear region and less frequently on the cell surface. After expression of exogenous AS160, GLUT4 on the cell surface decreased and GLUT4 vesicles were redistributed throughout the cytoplasm. Although PMA-induced or sodium fluoride-induced GLUT4 translocation was significantly increased in these cells, insulin did not affect GLUT4 translocation. These results suggest that AS160 is a common regulator of insulin- and GalphaqPCR activation-mediated GLUT4 distribution in the cells.
(キーワード): 3T3-L1 Cells / AMP-Activated Protein Kinases / Animals / CHO Cells / Cricetinae / Cricetulus / GTP-Binding Protein alpha Subunits, Gq-G11 / GTPase-Activating Proteins / Glucose Transporter Type 4 / Humans / Insulin / Mice / Rats
(出版サイトへのリンク): ● Publication site (DOI): 10.1507/endocrj.K08E-216
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19139597; ● Search Scopus @ Elsevier (PMID): 19139597; ● Search Scopus @ Elsevier (DOI): 10.1507/endocrj.K08E-216

(DOI: 10.1507/endocrj.K08E-216, PubMed: 19139597) Keiji Uchiyama, G Totskawa, M Puhka, Y Kaneko, E Jokitalo, I Dreveny, F Beuron, X Zhang, P Freemont and H Kondo :
p37is a p97 adaptor required for Golgi and ER biogenesis in interphase and at the end of mitosis.,
Developmental Cell, Vol.11, No.6, 803-816, 2006.

(要約): We previously reported that p97/p47-assisted membrane fusion is important for the reassembly of organelles at the end of mitosis, but not for their maintenance during interphase. We have now identified a p97 adaptor protein, p37, which forms a complex with p97 in the cytosol and localizes to the Golgi and ER. siRNA experiments revealed that p37 is required for Golgi and ER biogenesis. Injection of anti-p37 antibodies into cells at different cell cycle stages showed that p37 plays an important role in both Golgi and ER maintenance during interphase as well as in their reassembly at the end of mitosis. In an in vitro Golgi reassembly assay, the p97/p37 complex has membrane fusion activity. In contrast to the p97/p47 pathway, this pathway requires p115-GM130 tethering and SNARE GS15, but not syntaxin5. Interestingly, although VCIP135 is also required, its deubiquitinating activity is unnecessary for p97/p37-mediated activities.
(キーワード): ゴルジ体 / 小胞体 / p97 / p37
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.devcel.2006.10.016
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 17141156; ● Search Scopus @ Elsevier (PMID): 17141156; ● Search Scopus @ Elsevier (DOI): 10.1016/j.devcel.2006.10.016

(DOI: 10.1016/j.devcel.2006.10.016, PubMed: 17141156) F Kano, H Kondo, A Yamamoto, Y Kaneko, Keiji Uchiyama, N Hosokawa, K Nagata and M Murata :
NSF/SNAPs and p97/p47/VCIP135 are sequentially required for cell cycle-dependent reformation of the ER network.,
Genes to Cells, Vol.10, No.10, 989-999, 2005.

(要約): The endoplasmic reticulum (ER) has a characteristic polygonal structure with hallmark three-way junctions. In a previous paper, we reconstituted the disruption of the pre-existing ER network using mitotic cytosol from HeLa cells in streptolysin O (SLO)-permeabilized CHO-HSP cells (stably expressing GFP-HSP47). In addition, we found that interphase cytosol induced reformation of the disrupted ER network into a continuous network structure. Here, we show that the reformation of the ER network is accomplished through two sequential fusion reactions. The first process is mediated by NSF/alpha and gamma-SNAPs, and involves the generation of typical membranous intermediate structures that connect the disrupted ER tubules. A subsequent fusion is mediated by p97/p47/VCIP135, which has been shown to be required for homotypic fusion events in Golgi cisternae regrowth after mitosis. In addition, we also found that both fusion processes involve the t-SNARE, syntaxin 18.
(キーワード): ER network / NSF/SNAPs / p97/p47/VCIP135
(出版サイトへのリンク): ● Publication site (DOI): 10.1111/j.1365-2443.2005.00894.x
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16164599; ● Search Scopus @ Elsevier (PMID): 16164599; ● Search Scopus @ Elsevier (DOI): 10.1111/j.1365-2443.2005.00894.x

(DOI: 10.1111/j.1365-2443.2005.00894.x, PubMed: 16164599) X Yuan, P Simpson, C Mckeown, H Kondo, Keiji Uchiyama, R Wallis, I Dreveny, C Keetch, X Zhang, C Robinson, P Freemont and S Matthews :
Structure, dynamics and interactions of p47, a major adaptor of the AAA ATPase, p97.,
The EMBO Journal, Vol.23, No.7, 1463-1473, 2004.

(要約): p47 is a major adaptor molecule of the cytosolic AAA ATPase p97. The principal role of the p97-p47 complex is in regulation of membrane fusion events. Mono-ubiquitin recognition by p47 has also been shown to be crucial in the p97-p47-mediated Golgi membrane fusion events. Here, we describe the high-resolution solution structures of the N-terminal UBA domain and the central domain (SEP) from p47. The p47 UBA domain has the characteristic three-helix bundle fold and forms a highly stable complex with ubiquitin. We report the interaction surfaces of the two proteins and present a structure for the p47 UBA-ubiquitin complex. The p47 SEP domain adopts a novel fold with a betabetabetaalphaalphabeta secondary structure arrangement, where beta4 pairs in a parallel fashion to beta1. Based on biophysical studies, we demonstrate a clear propensity for the self-association of p47. Furthermore, p97 N binding abolishes p47 self-association, revealing the potential interaction surfaces for recognition of other domains within p97 or the substrate.
(キーワード): Adaptor Proteins, Signal Transducing / Adenosine Triphosphatases / Amino Acid Sequence / Animals / Humans / Models, Molecular / Molecular Sequence Data / Nuclear Magnetic Resonance, Biomolecular / Nuclear Proteins / Protein Folding / Protein Structure, Quaternary / Protein Structure, Secondary / Rats / Sequence Alignment / Ubiquitin
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/sj.emboj.7600152
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15029246; ● Search Scopus @ Elsevier (PMID): 15029246; ● Search Scopus @ Elsevier (DOI): 10.1038/sj.emboj.7600152

(DOI: 10.1038/sj.emboj.7600152, PubMed: 15029246) I Dreveny, H Kondo, Keiji Uchiyama, A Shaw, X Zhang and P Freemont :
Structural basis of the interaction between p97/VCP and its adaptor protein p47.,
The EMBO Journal, Vol.23, No.5, 1030-1039, 2004.

(要約): The AAA ATPase p97/VCP is involved in many cellular events including ubiquitin-dependent processes and membrane fusion. In the latter, the p97 adaptor protein p47 is of central importance. In order to provide insight into the molecular basis of p97 adaptor binding, we have determined the crystal structure of p97 ND1 domains complexed with p47 C-terminal domain at 2.9 A resolution. The structure reveals that the p47 ubiquitin regulatory X domain (UBX) domain interacts with the p97 N domain via a loop (S3/S4) that is highly conserved in UBX domains, but is absent in ubiquitin, which inserts into a hydrophobic pocket between the two p97 N subdomains. Deletion of this loop and point mutations in the loop significantly reduce p97 binding. This hydrophobic binding site is distinct from the predicted adaptor-binding site for the p97/VCP homologue N-ethylmaleimide sensitive factor (NSF). Together, our data suggest that UBX domains may act as general p97/VCP/CDC48 binding modules and that adaptor binding for NSF and p97 might involve different binding sites. We also propose a classification for ubiquitin-like domains containing or lacking a longer S3/S4 loop.
(キーワード): p97 / p47
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/sj.emboj.7600139
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 14988733; ● Search Scopus @ Elsevier (PMID): 14988733; ● Search Scopus @ Elsevier (DOI): 10.1038/sj.emboj.7600139

(DOI: 10.1038/sj.emboj.7600139, PubMed: 14988733) Keiji Uchiyama, Eija Jokitalo, Mervi Lindman, Mark Jackman, Fumi Kano, Masayuki Murata, Xiaodong Zhang and Hisao Kondo :
The localization and phosphorylation of p47 are important for Golgi disassembly-assembly during the cell cycle.,
The Journal of Cell Biology, Vol.161, No.6, 1067-1079, 2003.

(要約): In mammalian cells, the Golgi apparatus is disassembled at the onset of mitosis and reassembled at the end of mitosis. This disassembly-reassembly is generally believed to be essential for the equal partitioning of Golgi into two daughter cells. For Golgi disassembly, membrane fusion, which is mediated by NSF and p97, needs to be blocked. For the NSF pathway, the tethering of p115-GM130 is disrupted by the mitotic phosphorylation of GM130, resulting in the inhibition of NSF-mediated fusion. In contrast, the p97/p47 pathway does not require p115-GM130 tethering, and its mitotic inhibitory mechanism has been unclear. Now, we have found that p47, which mainly localizes to the nucleus during interphase, is phosphorylated on Serine-140 by Cdc2 at mitosis. The phosphorylated p47 does not bind to Golgi membranes. An in vitro assay shows that this phosphorylation is required for Golgi disassembly. Microinjection of p47(S140A), which is unable to be phosphorylated, allows the cell to keep Golgi stacks during mitosis and has no effect on the equal partitioning of Golgi into two daughter cells, suggesting that Golgi fragmentation-dispersion may not be obligatory for equal partitioning even in mammalian cells.
(キーワード): p97 / p47 / リン酸化 (phosphorylation) / ゴルジ体
(出版サイトへのリンク): ● Publication site (DOI): 10.1083/jcb.200303048
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12810701; ● Search Scopus @ Elsevier (PMID): 12810701; ● Search Scopus @ Elsevier (DOI): 10.1083/jcb.200303048

(DOI: 10.1083/jcb.200303048, PubMed: 12810701) Keiji Uchiyama, E Jokitalo, F Kano, M Murata, X Zhang, B Canas, R Newman, C Rabouille, D Pappin, P Freemont and H Kondo :
VCIP135, a novel essential factor for p97/p47-mediated membrane fusion, is required for Golgi and ER assembly in vivo.,
The Journal of Cell Biology, Vol.159, No.5, 855-866, 2002.

(要約): NSF and p97 are ATPases required for the heterotypic fusion of transport vesicles with their target membranes and the homotypic fusion of organelles. NSF uses ATP hydrolysis to dissociate NSF/SNAPs/SNAREs complexes, separating the v- and t-SNAREs, which are then primed for subsequent rounds of fusion. In contrast, p97 does not dissociate the p97/p47/SNARE complex even in the presence of ATP. Now we have identified a novel essential factor for p97/p47-mediated membrane fusion, named VCIP135 (valosin-containing protein [VCP][p97]/p47 complex-interacting protein, p135), and show that it binds to the p97/p47/syntaxin5 complex and dissociates it via p97 catalyzed ATP hydrolysis. In living cells, VCIP135 and p47 are shown to function in Golgi and ER assembly.
(キーワード): VCIP135 / p97 / p47 / 膜融合 / ゴルジ体 / 小胞体
(出版サイトへのリンク): ● Publication site (DOI): 10.1083/jcb.200208112
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12473691; ● Search Scopus @ Elsevier (PMID): 12473691; ● Search Scopus @ Elsevier (DOI): 10.1083/jcb.200208112

(DOI: 10.1083/jcb.200208112, PubMed: 12473691) S Abd-Aziz, C D Ang, M H Yusof, A M I Karim, A Ariff, Keiji Uchiyama and S Shioya :
Effect of C/N ratio and starch concentration on ethanol production from sago starch using recombinant yeast.,
World Journal of Microbiology & Biotechnology, Vol.17, 713-719, 2001. D Ang, S Abd-Aziz, M H Yusof, A M I Karim, A Ariff, Keiji Uchiyama and S Shioya :
Partial purification and characterization of amylolytic enzymes obtained from direct fermentation of Sago starch to ethanol by recombinant yeast,
Pakistan Journal of Biological Sciences, Vol.4, No.3, 266-270, 2001. C Lostainkit, Keiji Uchiyama, S Ochi, T Takaoka, K Nagahisa and S Shioya :
Characterization of bacteriocin N15 produced by Enterococcus faecium N15 and cloning of the related genes.,
Journal of Bioscience and Bioengineering, Vol.91, No.4, 390-395, 2001.

(要約): Enterococcus faecium N15 was isolated from nuka (Japanese rice-bran paste), which is utilized as starter in the fermenting of vegetables, and was found to produce a bacteriocin that exhibited a broad spectrum of activity, including activity against Listeria monocytogenes and Bacillus circulans JCM2504. The bacteriocin was sensitive to proteases (alpha-chymotrypsin, proteinase K, trypsin, and pepsin) and alpha-amylase, but it was resistant to lipase. The bacteriocin was resistant to heat treatment at 100 degrees C for 2 h, but its activity was completely lost after autoclaving at 121 degrees C for 15 min. It was active over a wide pH range from 2.0 to 10.0. The bacteriocin showed bactericidal activity against Lactobacillus sake JCM1157 at a concentration of 40 AU/ml. Its molecular weight was estimated by SDS-PAGE to be about 3-5 kDa. PCR primers were designed based on the conserved amino acid sequences of class IIa bacteriocins. A 3-kb DNA fragment was amplified and three open reading frames (ORFs) were found. The first encodes a probable immunity protein of 103 amino acid residues and shows complete homology with the putative immunity protein of E. faecium DPC1146. The second and third ORFs respectively encode a probable transposase gene and an inducing factor. The upstream region of the immunity gene, in which the bacteriocin structural gene is located, was amplified. A homology search revealed that the bacteriocin produced by E. faecium N15 exhibits complete identity to enterocin A, a bacteriocin produced by E. faecium DPC1146. PCR using the primers designed in this study is a rapid and sufficient method of screening for bacteriocin-producing strains.
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/S1389-1723(01)80157-2
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16233010; ● Search Scopus @ Elsevier (PMID): 16233010; ● Search Scopus @ Elsevier (DOI): 10.1016/S1389-1723(01)80157-2

(DOI: 10.1016/S1389-1723(01)80157-2, PubMed: 16233010) Keiji Uchiyama and S Shioya :
Modeling and optimization of alpha-amylase production in a recombinant yeast fed-batch culture taking account of the cell cycle population distribution.,
Journal of Biotechnology, Vol.71, 133-141, 1999. H D Lee, Keiji Uchiyama, S Shioya and I Y Hwang :
Maximum glucoamylase production by a temperature-sensitive mutant of Saccharomyces cerevisiae in batch culture.,
Journal of Industrial Microbiology & Biotechnology, Vol.20, 160-165, 1998. H Shimizu, K Yasuoka, Keiji Uchiyama and S Shioya :
Bioprocess fault detection by nonlinear multivariate analysis: application of an artificial autoassociative neural network and wavelet filter bank.,
Biotechnology Progress, Vol.14, No.1, 79-87, 1998. H Shimizu, K Yasuoka, Keiji Uchiyama and S Shioya :
On-line fault diagnosis for an optimal rice -amylase production process of temperature-sensitive mutant of Saccharomyces cerevisiae by autoassociative neural network.,
Journal of Fermentation and Bioengineering, Vol.83, No.5, 435-442, 1997. Keiji Uchiyama, M Morimoto, Y Yokoyama and S Shioya :
Cell cycle dependency of rice alpha-amylase production in a recombinant yeast.,
Biotechnology and Bioengineering, Vol.54, No.3, 262-271, 1997. Keiji Uchiyama, T Ohotani, M Morimoto, S Shioya, K Suga, S Harashima and Y Oshima :
Optimization of rice alpha-amylase production using temperature-sensitive mutants of Saccharomyces cerevisiae for the PHO regulatory system.,
Biotechnology Progress, Vol.11, No.5, 510-517, 1995.

MISC

研究者総覧に該当データはありませんでした。

総説・解説

内山圭司, 坂口末廣 :
プリオン病における神経変性のメカニズム,
Clinical Neuroscience, Vol.31, No.9, 1022-1024, 2013年9月. 近藤久雄, 内山圭司, 十津川剛 :
細胞内小器官の維持のための新たな分子機構,
実験医学, Vol.25, No.6, 848-851, 2007年4月.

(キーワード): ゴルジ体 / 小胞体 (endoplasmic reticulum) / p97 / p37 / 細胞内膜融合

近藤久雄, 内山圭司, 十津川剛 :
細胞内小器官の維持のための新たな分子機構,
実験医学, Vol.25, No.6, 848-851, 2007年4月.

(キーワード): 細胞内小器官 / ゴルジ体 / 細胞内膜融合

Keiji Uchiyama and H Kondo :
p97/p47-Mediated Biogenesis of Golgi and ER.,
The Journal of Biochemistry, Vol.137, No.2, 115-119, Feb. 2005.

(要約): In mammalian cells, the Golgi apparatus and endoplasmic reticulum have typical structures during interphase: stacked cisternae located adjacent to the nucleus and a network of interconnected tubules throughout the cytoplasm, respectively. At mitosis their architectures disappear and are reassembled in daughter cells. p97, an AAA-ATPase, mediates membrane fusion and is required for reassembly of these organelles. In the p97-mediated membrane fusion, p47 was identified as an essential cofactor, through which p97 binds to a SNARE, syntaxin5. A second essential cofactor, VCIP135, was identified as a p97/p47/syntaxin5-interacting protein. Several lines of recent evidence suggest that ubiquitination may be implicated in the p97/p47 pathway; p47 binds to monoubiquitinated proteins and VCIP135 shows a deubiquitinating activity in vitro. For the cell-cycle regulation of the p97/p47 pathway, it has been reported that the localization and phosphorylation-dephosphorylation of p47 are crucial. In this review, we describe the components involved in the p97-mediated membrane fusion and discuss the regulation of the fusion pathway.
(キーワード): p97 / p47 / ゴルジ体 / 小胞体
(出版サイトへのリンク): ● Publication site (DOI): 10.1093/jb/mvi028
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15749824; ● Search Scopus @ Elsevier (PMID): 15749824; ● Search Scopus @ Elsevier (DOI): 10.1093/jb/mvi028

(DOI: 10.1093/jb/mvi028, PubMed: 15749824) 近藤久雄, 内山圭司, 沖米田司 :
ゴルジ体・小胞体形成に必須なp97膜融合機構,
蛋白質核酸酵素, Vol.49, No.7, 916-919, 2004年5月.

(キーワード): Adenosine Triphosphatases / Carrier Proteins / Cell Cycle / Endopeptidases / Endoplasmic Reticulum / Golgi Apparatus / Membrane Fusion / Membrane Proteins / Nuclear Proteins / SNARE Proteins / Vesicular Transport Proteins
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15168492; ● Search Scopus @ Elsevier (PMID): 15168492

(PubMed: 15168492) 内山圭司 :
環境修復技術としてのコンポスト化,
生物工学会誌, Vol.78, No.2, 67, 2000年. 塩谷捨明, 清水浩, 内山圭司 :
バクテリオシン産生乳酸菌の培養工学,
生物工学会誌, Vol.77, No.11, 461-462, 1999年. 内山圭司, 塩谷捨明 :
生分解性プラスチックのコンポスト化技術,
化学装置, Vol.40, 94-98, 1998年7月. 塩谷捨明, 清水浩, 内山圭司 :
乳酸菌によるバクテリオシンの培養工学,
生物工学会誌, Vol.76, No.11, 460-461, 1998年.

講演・発表

K Kaneyoshi, K Kuroda, N Yamano, Y Koga, Keiji Uchiyama and Takeshi Omasa :
Analysis of intracellular secretion processes by Citrine fusion IgG aiming to establish high producer CHO cells. Analysis of intracellular secretion processes by Citrine fusion IgG,
JAACT2018 Tsukuba, Tsukuba, Nov. 2018. K Kaneyoshi, Keiji Uchiyama, Masayoshi Onitsuka, Noriko Yamano, Y Koga and Takeshi Omasa :
Intracellular secretion analysis of recombinant therapeutic antibodies in engineered CHO cells aiming to establish high produce.,
The 25th Meeting of the European Society for Animal Cell Technology (ESACT2017), May 2017. Kohei Kaneyoshi, Keiji Uchiyama, Masayoshi Onitsuka, Noriko Yamano, Yuichi Koga and Takeshi Omasa :
Analysis of intracellular recombinant IgG secretion in engineered CHO cells,
The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2016 Kobe), Nov. 2016. Keiji Uchiyama and Suehiro Sakaguchi :
Sorting of prion protein and PrPSc accumulation.,
International Prion Congress-Prion 2016, Tokyo, May 2016. Keiji Uchiyama, Tomita Mitsuru and Suehiro Sakaguchi :
Mechanism of sortilin-mediated PrP degradation.,
Asian Pacific Prion Symposium 2015, Ishikawa Ongakudo, Kanazawa, Japan., Sep. 2015. Tomita Mitsuru, Keiji Uchiyama, Usui Takeshi and Suehiro Sakaguchi :
Analysis of roles of N-terminal region of prion protein in the conversion into scrapie form by using prion-knockout cell.,
Asian Pacific Prion Symposium 2015, Ishikawa Ongakudo, Kanazawa, Japan, Sep. 2015. Keiji Uchiyama and Suehiro Sakaguchi :
Post-Golgi trafficking of membrane proteins impaired by prion infection.,
Prion, Vol.8, 98, Trieste, Italy, Jun. 2014.

(キーワード): prion / membrane trafficking

Keiji Uchiyama and Suehiro Sakaguchi :
Prions disturb post-Golgi membrane trafficking to the cell surface.,
Proceedings of The 9th International Symposium of the Institute Network, 30, Osaka, Jun. 2014. Suehiro Sakaguchi and Keiji Uchiyama :
Prions impair post-Golgi trafficking of membrane proteins,
Asian Pacific Prion Symposium 2013, Nagasaki, Jul. 2013. Keiji Uchiyama, G Totsukawa, M Rantanen, E Jokitalo and H Kondo :
p97-mediated biogenesis of the Golgi and ER.,
20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress, Kyoto, Jun. 2006. Keiji Uchiyama, G Totsukawa and H Kondo :
p37, a novel adaptor of p97, is required for Golgi and ER maintenance during interphase.,
International Symposium on Life of Proteins, Hyogo, Oct. 2005. Keiji Uchiyama, T Suzuki, H Tatsumi, H Kanetake and S Shioya :
Amplified 16S Ribosomal DNA Restriction Analysis of Microbial Community Structure During Fast Degradation of a Biopolymer, PHA, by Composting.,
Proceeding of the International Symposium on Microbiology of Composting, Innsbruck, Oct. 2000. S Shioya, H Kodama, T Arimoto, Keiji Uchiyama and H Shimizu :
New Process Development for Mixed Culture Experiments,
Proceedings of the 5th Asia-Pacific Biochemical Engineering Conference, O-BP1, Nov. 1999. Keiji Uchiyama, C Losteinkit, S Ochi and S Shioya :
Isolation and characterization of bacteriocin-producing Enterococcus faecium N15 from Japanese fermented food.,
Proceeding of Sixth Symposium on Lactic Acid Bacteria, C12, Veldhoven, Netherlands, Sep. 1999. Keiji Uchiyama and S Shioya :
Modeling of rice -amylase production process in a recombinant yeast taking account of the cell cycle population.,
Proceeding of the 7th International Conference on Computer Application in Biotechnology, Osaka, May 1999. Keiji Uchiyama, T Suzuki, H Tatsumi and S Shioya :
Microbial population analysis during composting process using ARDRA.,
Proceeding of 8th International Symposium on Microbial Ecology., Halifax, Canada, Sep. 1998. H Shimizu, K Yasuoka, Keiji Uchiyama and S Shioya :
On-line Fault Detection for an Optimal Rice alpha-amylase Production Process of Saccharomyces cerevisiae by an Autoassociative Neural Network.,
Proceeding of Engineering Foundation, Chemical Engineering X, 4-6, May 1997. S Shioya, Keiji Uchiyama, M Morimoto, K Suga, S Harashima and Y Oshima :
Cell cycle dependent rice alpha-amylase production by a recombinant yeast.,
the 9th NCRT; DOST-JSPS joint seminar on Biotechnology, Khon Kaen, Thailand, Oct. 1994. S Shioya, Keiji Uchiyama, T Ohtani, A Kanda, K Suga, S Harashima and Y Oshima :
Maximum production in batch culture of S. cerevisiae using temperature-dependent gene expression system.,
Better living through innovative biochemical engineering, 708-710, 1994. S Shioya, Keiji Uchiyama, T Ohtani, A Kanda, K Suga, S Harashima and Y Oshima :
Maximum production in batch culture of S. cerevisiae using temperature-dependent gene expression system.,
Proceeding of Seminar on Biochemical Engineering, 35-37, Taipei, Taiwan ROC, Dec. 1993. S Shioya, Keiji Uchiyama, T Ohtani, A Kanda, K Suga, S Harashima and Y Oshima :
Maximum production in batch culture of S. cerevisiae using temperature-dependent gene expression system.,
Proceeding of 12th IFAC World Congress, Vol.8, 231-236, Sydney, Jul. 1993. S Shioya, Keiji Uchiyama, T Ohtani, A Kanda, K Suga, S Harashima and Y Oshima :
Modeling and control for maximum production of S. cerevisiae using temperature-dependent gene expression system.,
Proceeding of 6th ECB, Vol.II, TU-390, Firenze, Jun. 1993. 内山圭司, 吉丸哲郎, 松下洋輔, 尾野雅哉, 三好康雄, 笹三徳, 片桐豊雅 :
乳がん細胞における O型糖鎖修飾を介した IRE1の恒常的活性化機構,
第95回日本生化学会大会, 2022年11月. 萩原浩生, 内山圭司, 吉丸哲郎, 松下洋輔, 三好康雄, 笹三徳, 片桐豊雅 :
乳がん細胞におけるO 結合型糖鎖修飾GRP78/Bip の役割解明と創薬研究,
第81回日本癌学会学術総会, 2022年10月. 内山圭司, 吉丸哲郎, 松下洋輔, 尾野雅哉, 笹三徳, 三好康雄, 片桐豊雅 :
乳がん細胞におけるO 結合型糖転移酵素の発現亢進はIRE1 の恒常的活性化を惹起する,
第81回日本癌学会学術総会, 2022年9月. 萩原浩生, 内山圭司, 吉丸哲郎, 松下洋輔, 片桐豊雅 :
乳がん細胞におけるO結合型糖鎖修飾GRP78/Bipの役割解明と創薬研究,
第26回日本がん分子標的治療学会学術集会, 2022年6月. 内山圭司, 松下洋輔, 吉丸哲郎, 萩原浩生, 片桐豊雅 :
乳がん細胞におけるO型糖鎖修飾IRE1経路の恒常的活性化を標的とした新規治療戦略の可能性,
第26回日本がん分子標的治療学会学術集会, 2022年6月. 兼吉航平, 黒田昂輝, 山野-足立範子, 鬼塚正義, 古賀雄一, 内山圭司, 大政健史 :
生産性向上を目指した医薬品抗体生産細胞における分泌過程解析,
日本化学工学会第84年会, G223, 2019年3月. 内山圭司, 坂口末廣 :
Identification and investigation of a novel anti-prion compound.,
第65回日本ウイルス学会学術集会, 2017年10月. 内山圭司, 藤稿智宏, 坂口末廣 :
High susceptibility of Sortilin-deficient cells to prion infection.,
第65回日本ウイルス学会学術集会, 2017年10月. 兼吉航平, 内山圭司, 鬼塚正義, 山野範子, 古賀雄一, 大政健史 :
高生産株の構築を目指した抗体生産CHO細胞内の分泌過程解,
第69回日本生物工学会大会, 2017年9月. 坂口末廣, 内山圭司 :
Prion propagation through sortilin degradation.,
第60回日本神経化学会大会, 2017年9月. 内山圭司, 坂口末廣 :
プリオン病における異常プリオンの蓄積メカニズム,
第39回日本分子生物学会, 2P-0593, 2016年12月. 内山圭司 :
Dysfunction of Sortilin by prion infection,
第64回日本ウイルス学会学術集会, P1-105, 2016年10月. 内山圭司, 富田満, 坂口末廣 :
プリオン感染により過剰な異常プリオンが蓄積する分子メカニズム,
第38回に本分子生物学会年会第88回日本生化学会大会合同大会, 2015年12月. 内山圭司, 富田満, 坂口末廣 :
Inhibition of Sortilin-mediated PrP degradation by prion infection causes excessive accumulation of abnormal prion protein.,
第63回日本ウイルス学会学術集会, 2015年11月. Keiji Uchiyama, 富田満 and Suehiro Sakaguchi :
Novel molecular mechanism for accumulation of abnormal prion protein - Inhibition of Sortilin-mediated PrP degradation.,
第15回蛋白質科学会年会ワークショップ「New development of prion biology」, Jun. 2015.

(キーワード): prion / Sortilin / degradation

坂口末廣, 内山圭司 :
プリオンによるポストゴルジ膜輸送障害,
第36回生体膜と薬物の相互作用シンポジウム「生体膜における蛋白質の機能制御システムと疾患」, 2014年11月. 富田満, 内山圭司, 臼井健, 坂口末廣 :
プリオンノックアウト細胞を用いたプリオンタンパク質N末端領域の異常プリオン形成における役割の解明,
第62回日本ウイルス学会学術集会, 2014年11月. 内山圭司, 富田満, 臼井健, 坂口末廣 :
新規プリオン結合因子Sortilinのプリオン感染における役割,
第62回日本ウイルス学会学術集会, 2014年11月. 内山圭司, 坂口末廣 :
プリオン感染と小胞輸送障害,
第87回日本生化学会シンポジウム認知症克服に向けて:プリオン病をもっと知る., 2014年10月. 内山圭司, 坂口末廣 :
プリオン感染によるSortilin発現低下が異常プリオン蓄積を引き起こす,
第29回中国四国ウイルス研究会プログラム・抄録集, 26, 2014年6月.

(キーワード): プリオン / 異常プリオン

内山圭司, 坂口末廣 :
プリオン感染によるポストゴルジ小胞輸送障害,
第66回日本細胞生物学会要旨集, 149, 2014年6月.

(キーワード): プリオン / 小胞輸送

内山圭司 :
神経変性疾患と小胞輸送-プリオン病における小胞輸送障害,
尚醸会バイオテクノロジーセミナー, 2013年11月.

(キーワード): 神経変性疾患 / 小胞輸送 / プリオン病

内山圭司, 坂口末廣 :
プリオン感染はポストゴルジ小胞輸送を抑制する,
第61回日本ウイルス学会学術集会, 2013年11月. 内山圭司, 坂口末廣 :
プリオン感染はポストゴルジ小胞輸送障害を引き起こす,
日本生化学会, 2013年9月. 松村しずか, 井川翔太, 鬼塚正義, 白井昭博, 間世田英明, 内山圭司, 大政健史 :
CHO細胞における小胞出芽関連因子Arfのクローニングと発現抑制によるタンパク質生産への影響,
日本動物細胞工学会2013年度大会(JAACT2013), 69, 2013年7月. 坂口末廣, 内山圭司 :
Inhibition of Post-Golgi Membrane Traffic in Prion-Infected Cells,
第35回分子生物学会年会(ポスター発表), 2012年12月. 坂口末廣, 内山圭司 :
Inhibition of Post-Golgi Membrane Traffic in Prion-Infected Cells,
第35回分子生物学会年会ワークショップ「Membrane Traccic and Diseases」(口頭発表), 2012年12月. 十津川剛, 内山圭司, 金子弥生, 小川洋子, 近藤久雄 :
p97-ATPase-mediated membrane fusion.,
第28回日本分子生物学会年会ワークショップ「細胞機能のキープレーヤーAAA+タンパク質の世界」, 2005年12月. 内山圭司, 近藤久雄 :
p97-mediated membrane fusion.,
第27回日本分子生物学会年会, ワークショップ「AAA+タンパク質の細胞機能と構造」, 2004年12月. 内山圭司, 加納ふみ, E Jokitlo, X Zhaug, 村田昌之, R Newman, D Pappin, 塩谷捨明, P Freemont, 近藤久雄 :
The p97/p47 pathway in Golgi reassembly.,
第24回日本分子生物学会年会, シンポジウム「細胞内メンブレントラフィックーOrganellar Identity and Dynamics―」, 2001年12月. Keiji Uchiyama, M Morimoto, S Shioya, K Suga, S Harashima and Y Oshima :
Rice alpha-amylase production by a recombinant yeast.,
第27回シンポジウムシリーズ, 110-115, 1994.

研究会・報告書

内山圭司 :
プリオン病-異常プリオン蓄積メカニズムの解明,
「核・ミトコンドリア遺伝子の網羅的トランスゲノム解析による心不全重症化機構の解明」研究班シンポジウム「蛋白質の構造と病態」, 2016年2月.

(キーワード): プリオン病 / 異常プリオン / タンパク質分解 / 細胞内小胞輸送

内山圭司, 臼井健, 矢野雅司, 坂口末廣 :
プリオン感染はポストゴルジ小胞輸送を抑制する,
第28回中国四国ウイルス研究会, 2013年6月. 内山圭司, 臼井健, 矢野雅司, 坂口末廣 :
プリオン感染によるポストゴルジ小胞輸送抑制,
第54回日本生化学会中国・四国支部例会, 2013年6月. 内山圭司, 坂口末廣 :
プリオン感染は神経細胞におけるインスリンシグナル異常を引き起こす,
第27回中国四国ウイルス研究会抄録集, 2012年6月.

(キーワード): プリオン

内山圭司, 蛯名洋介 :
Functional roles of TUG and p97/VCP in Glut4 trafficking.,
第34回分子生物学会年会, 4T14a-3, 2011年12月.

(キーワード): Glut4 / TUG / p97 / VCP

内山圭司, 蛯名洋介 :
Functional roles of TUG and p97/VCP in Glut4 trafficking.,
第34回分子生物学会年会, 3P-0673, 2011年12月.

(キーワード): Glut4 / TUG / p97 / VCP

内山圭司, 坂口末廣 :
プリオン感染細胞における細胞内小胞輸送の抑制,
第26回中国四国ウイルス研究会抄録集, 2011年6月.

(キーワード): プリオン / ゴルジ体

坂口末廣, 宮田博規, 山口仁孝, 村松直美, 森剛志, 内山圭司, 犬伏祥子 :
異なるプリオン株の産生メカニズムについて,
第26回中国四国ウイルス研究会徳島大学青藍会館徳島, 2011年6月. Keiji Uchiyama and Yousuke Ebina :
Role of BIG2, a Guanine-Nucleotide Exchanging Factor for ADP-Ribosylation Factors, in Insulin-Regulated Glucose Transporter Translocation,
第32回日本分子生物学会年会, 168, Dec. 2009.

特許: 塩谷捨明, 清水浩, 内山圭司 : 分離型混合培養装置, 特願981505, .
作品: 研究者総覧に該当データはありませんでした。
補助金・競争的資金: がん微小環境でのIRE1恒常的活性化に必須な小胞体-ゴルジ体間シャトル輸送機構の解明 (研究課題/領域番号: 23K06651 )
BIG3-PHB2複合体を標的としたHER2陽性乳がんの薬剤耐性を克服する多角的がん治療戦略 (研究課題/領域番号: 23H02970 )
細胞内小胞輸送活性化による老化脳タンパク質分解促進機構の解明と治療への応用 (研究課題/領域番号: 17H05702 )
プリオン感染によるソーティリン発現量低下機構の解明と治療法開発への応用 (研究課題/領域番号: 17K08859 )
新規プリオン結合因子ソーティリンによるプリオン病治療基盤技術の開発 (研究課題/領域番号: 26460557 )
Glut4トランスロケーショに関する新規必須因子BIG2の機能解析 (研究課題/領域番号: 21570142 )
インスリンシグナル伝達の分子機構と糖尿病 (研究課題/領域番号: 20390095 )
p97/p37新規膜融合経路に関する研究-SNARE複合体の同定と機能解析 (研究課題/領域番号: 19570195 )
インスリンシグナル伝達の分子機構と糖尿病 (研究課題/領域番号: 18390104 )
新規の細胞内膜融合機構p97/p37経路の解明 (研究課題/領域番号: 18050046 )
p97を介したゴルジ体再構成過程におけるVCIP135による脱ユビキチン化の役割 (研究課題/領域番号: 17770180 )
小胞体の網状構造の形成機構 (研究課題/領域番号: 17028060 )
抗菌物質ナイシン生産プロセスにおける微生物相互作用を利用した制御技術の開発 (研究課題/領域番号: 10650784 )
乳酸菌による新規バクテリオシンの生産とその食品保存料への展開 (研究課題/領域番号: 10450311 )
温度感受性組換え酵母培養プロセスにおけるオンライン状態認識・制御システムの開発 (研究課題/領域番号: 08455381 )
研究者番号（60294039）による検索

その他: 研究者総覧に該当データはありませんでした。

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2024年4月25日更新

専門分野・研究分野: 分子細胞生物学 (Molecular and Cellular Biology)
神経化学 (Neurochemistry)
分子細胞生物学 (Molecular and Cellular Biology)
所属学会・所属協会: 日本分子生物学会
日本細胞生物学会
日本生化学会
日本ウイルス学会
委員歴・役員歴: 日本ウイルス学会 ( [2013年]) (アジア太平洋プリオン研究会 [2016年10月])
受賞: 2014年1月, 康楽賞 (財団法人三木康楽会)
2015年9月, Asia Pacific Prion Symposium 2015, The Best Poster Award (Asia Pacific Society of Prion Research (APSPR))
2016年11月, Best Oral Presentation Awards (The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology)
活動: 毒物・劇物管理担当 (2011年4月〜2021年3月)
環境・エネルギー管理委員会委員 (2012年4月〜2016年3月)
研究所セミナー運営委員 (2016年4月〜2019年12月)
蔵本地区駐車場委員会委員 (2018年4月〜2020年3月)
先端酵素学研究所放射線安全管理委員会 (2019年11月〜2021年3月)
蔵本地区駐車場委員会委員 (2020年4月〜2021年3月)

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2024年4月21日更新

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Ｊグローバル

Jグローバル最終確認日: 2024/4/20 01:18
氏名（漢字）: 内山圭司
氏名（フリガナ）: ウチヤマケイジ
氏名（英字）: KEIJI UCHIYAMA
所属機関: 徳島大学先端酵素学研究所特任准教授

リサーチマップ

researchmap最終確認日: 2024/4/21 01:24
氏名（漢字）: 内山圭司
氏名（フリガナ）: ウチヤマケイジ
氏名（英字）: KEIJI UCHIYAMA
プロフィール: リサーチマップAPIで取得できませんでした。
登録日時: 2018/11/14 08:50
更新日時: 2024/2/2 03:07
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所属ID: 0344000000
所属: 徳島大学先端酵素学研究所
部署: ゲノム制御学分野
職名: 特任准教授
学位: 工学博士
学位授与機関: 大阪大学
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ＫＡＫＥＮで最新データを確認する

2024年4月20日更新

研究者番号: 60294039

所属（現在）: 2024/4/1 : 徳島大学, 先端酵素学研究所, 特任准教授

所属（過去の研究課題 情報に基づく）*注記: 2023/4/1 : 徳島大学, 先端酵素学研究所, 特任准教授
2016/4/1 – 2019/4/1 : 徳島大学, 先端酵素学研究所(次世代), 准教授
2014/4/1 – 2015/4/1 : 徳島大学, 疾患酵素学研究センター, 准教授
2007/4/1 – 2011/4/1 : 徳島大学, 疾患酵素学研究センター, 准教授
2006/4/1 : 三菱化学生命科学研究所, 研究部門, 研究員
2005/4/1 – 2006/4/1 : 株式会社三菱化学生命科学研究所, 研究部門, 研究員
2005/4/1 : 株式会社三菱化学生命科学研究所, 研究員
1998/4/1 – 1999/4/1 : 大阪大学, 大学院・工学研究科, 助手
1997/4/1 : 大阪大学, 工学部, 助手

審査区分/研究分野

研究代表者

生物系 / 生物学 / 生物科学 / 細胞生物学
生物系 / 生物学 / 生物科学 / 機能生物化学
生物系 / 医歯薬学 / 基礎医学 / ウイルス学
生物系
小区分50010:腫瘍生物学関連

研究代表者以外

生物系 / 医歯薬学 / 基礎医学 / 病態医化学
工学 / プロセス工学 / 生物・生体工学
生物系
小区分55010:外科学一般および小児外科学関連

キーワード

研究代表者

ゴルジ体 / p97 / VCIP135 / ユビキチン / 膜融合 / 97 / VCP / VCP135 / p47 / 細胞構造・機能 / 細胞内膜融合 / SNARE / p37 / Glut4 / BIG2 / Glut4トランスロケーション / インスリン作用 / 脂肪細胞 / インスリン / Glut4小胞再構成 / リサイクリングエンドソーム / 3T3-L1脂肪 / プリオン / プリオン病 / 異常プリオン / Sortilin / ソーティング / レトロマー / VPS35 / ソーティリン / プリオン感染 / レトロマー複合体 / タンパク質分解 / プリオン分解 / タパク質分解 / プリオン高感受性 / ンソーティリン欠損マウス / ソーティリン欠損細胞 / 抗プリオン活性 / 神経変性疾患 / 抗プリン活性 / 細胞内小胞輸送 / エタノールアミン / T0901317 / Liver X receptor / がん微小環境 / 小胞体ストレス / O型糖鎖修飾

研究代表者以外

小胞体 / ゴルジ体 / 膜融合 / p97ATPase / p37 / p47 / 網状構造 / VCIP135 / 分子病態学 / インスリンシグナル伝達 / インスリン作用 / GLUT4 / TUG / sIR / 高血糖 / 超高感度sIR測定法 / シグナル伝達 / BFA / GLUT4トランスロケーション / ARF / BIG2 / 温度感受性組換え酵母 / 異常診断 / 自己想起型ニューラルネットワーク / ウェーブレットフィルタ / 状態認識 / ウェーブレット変換 / temperature controllable gene expression / autoassociative neural network / fault diagnosis / wavelet filter / エンテロシンA / バクテリオシン / シャトルベクター / E.faecium / バクテリオシン耐性遺伝子 / 発酵食品 / 乳酸菌 / Lactobacillus plantarum / Enterococcus faecium / enterocin A / bacteriocin / shuttle vector / E. faecium / immunity for enterocin A / 酵母 / 混合培養 / ナイシン / カスケード制御 / 溶存酸素濃度 / マルトース / ラクトース / lactic acid bacteria / yeasts / mixed culture / nisin / cascade controller / dissolved oxygen concentration / maltose / インスリン受容体 / 可溶性細胞外ドメイン / 糖尿病 / AS160 / Gq共役受容体 / インスリン / insulin receptor / soluble ectodomain / diabetes / signal transduction / GLUT4 translocation / Gq coupled receptor / HER2陽性乳がん / トラスツズマブ耐性 / ペプチド創薬 / がん抑制因子 / ステープル化ペプチド

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