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ニーマン・ピックC1の小胞体関連分解熱ショック蛋白質の役割の証明とユビキチンを受け入れるリジン残基の同定
https://repository.lib.tottori-u.ac.jp/records/5844
https://repository.lib.tottori-u.ac.jp/records/584431814c9e-75ce-4942-b5e2-2fcd73806ecb
| 名前 / ファイル | ライセンス | アクション |
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本文 : Full Text (2.4 MB)
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要旨 : Summary (521.8 kB)
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審査結果要旨 : Summary of Thesis Review (493.0 kB)
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| Item type | 学位論文 / Thesis or Dissertation(1) | |||||
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| 公開日 | 2018-06-22 | |||||
| タイトル | ||||||
| タイトル | ニーマン・ピックC1の小胞体関連分解熱ショック蛋白質の役割の証明とユビキチンを受け入れるリジン残基の同定 | |||||
| 言語 | ja | |||||
| タイトル | ||||||
| タイトル | Endoplasmic reticulum-associated degradation of Niemann-Pick C1 : evidence for the role of heat shock proteins and identification of lysine residues that accept ubiquitin | |||||
| 言語 | en | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ | doctoral thesis | |||||
| アクセス権 | ||||||
| アクセス権 | open access | |||||
| 著者 |
仲宗根, 眞恵
× 仲宗根, 眞恵× Nakasone, Naoe |
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| 抄録 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | Most cases with Niemann-Pick disease type C carry mutations in NPC1. Some of the mutations, including the most frequent I1061T, give rise to unstable proteins selected for endoplasmic reticulum-associated degradation. The purpose of the current study was to shed mechanistic insights into the degradation process. A proteasome inhibitor MG132 prolonged the life span of the wild-type NPC1 expressed in COS cells. The expressed protein associated with multiple chaperones including heat shock protein 90 (Hsp90), Hsp70, heat shock cognate protein 70 (Hsc70), and calnexin. Accordingly, expression of an E3 ligase CHIP (carboxyl terminus of Hsp70-interacting protein) enhanced MG132-induced accumulation of ubiquitylated NPC1. Co-expression and RNAi knockdown experiments in HEK cells indicated that Hsp70/Hsp90 stabilized NPC1, whereas Hsc70 destabilized it. In human fibroblasts carrying the I1061T mutation, adenovirus-mediated expression of Hsp70 or treatment with an HSP-inducer geranylgeranylacetone (GGA) increased the level of the mutant protein. In GGA-treated cells, the rescued protein was localized in the late endosome and ameliorated cholesterol accumulation. MALDI-TOF mass spectrometry revealed three lysine residues at amino acids 318, 792, and 1180 as potential ubiquitin-conjugation sites. Substitutions of the three residues with alanine yielded a mutant protein with a steady-state level more than three times higher than that of the wild-type. Introduction of the same substitutions to the I1061T mutant resulted in an increase in its protein level and functional restoration. These findings indicated the role of HSPs in quality control of NPC1 and revealed the role of three lysine residues as ubiquitin-conjugation sites. | |||||
| 学位名 | ||||||
| 言語 | ja | |||||
| 学位名 | 博士(医学) | |||||
| 学位授与機関 | ||||||
| 学位授与機関識別子Scheme | kakenhi | |||||
| 学位授与機関識別子 | 15101 | |||||
| 言語 | ja | |||||
| 学位授与機関名 | 鳥取大学 | |||||
| 学位授与年月日 | ||||||
| 学位授与年月日 | 2017-09-30 | |||||
| 学位授与番号 | ||||||
| 学位授与番号 | 甲第860号 | |||||
| 権利 | ||||||
| 権利情報 | © the American Society for Biochemistry and Molecular Biology | |||||
| 情報源 | ||||||
| 関連名称 | This research was originally published in the Journal of Biological Chemistry. Naoe Nakasone, Yuko S. Nakamura, Katsumi Higaki, Nao Oumi, Kousaku Ohno and Haruaki Ninomiya. Endoplasmic reticulum-associated degradation of Niemann-Pick C1 evidence for the r | |||||
| 関連サイト | ||||||
| 識別子タイプ | URI | |||||
| 関連識別子 | http://www.jbc.org/content/289/28/19714 | |||||
| 関連名称 | http://www.jbc.org/content/289/28/19714 | |||||
| 著者版フラグ | ||||||
| 出版タイプ | VoR | |||||
