論文 - 渡辺 洋平
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The fission yeast Greatwall-Endosulfine pathway is required for proper quiescence/G0 phase entry and maintenance. 査読あり
Aono Soma, Haruna Yui, Watanabe Yo-hei, Mochida Satoru, Takeda Kojiro
Genes to Cells 24 ( 2 ) 172 - 186 2019年2月
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DOI： 10.1111/gtc.12665.
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Sugita Saori, Watanabe Kumiko, Hashimoto Kana, Niwa Tatsuya, Uemura Eri, Taguchi Hideki, Watanabe Yo-hei
293 ( 50 ) 19228 - 19239 2018年12月
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担当区分：最終著者, 責任著者
ClpB, a bacterial homologue of heat shock protein 104 (Hsp104), can disentangle aggregated proteins with the help of the DnaK, a bacterial Hsp70, and its co-factors. As a member of the expanded superfamily of ATPases associated with diverse cellular activities (AAA+), ClpB forms a hexameric ring structure, with each protomer containing two AAA+ modules, AAA1 and AAA2. A long coiled-coil middle domain (MD) is present in the C-terminal region of the AAA1 and surrounds the main body of the ring. The MD is subdivided into two oppositely directed short coiled-coils, called motif-1 and motif-2. The MD represses the ATPase activity of ClpB, and this repression is reversed by the binding of DnaK to motif-2. To better understand how the MD regulates ClpB activity, here we investigated the roles of motif-1 in ClpB from Thermus thermophilus (TClpB). Using systematic alanine substitution of the conserved charged residues, we identified functionally important residues in motif-1, and using a photoreactive cross-linker and LC-MS/MS analysis, we further explored potential interacting residues. Moreover, we constructed TClpB mutants in which functionally important residues in motif-1 and in other candidate regions were substituted by oppositely charged residues. These analyses revealed that the intra-subunit pair Glu-401-Arg-532 and the inter-subunit pair Asp-404-Arg-180 are functionally important, electrostatically interacting pairs. Considering these structural findings, we conclude that the Glu-401-Arg-532 interaction shifts the equilibrium of the MD conformation to stabilize the activated form and that the Arg-180-Asp-404 interaction contributes to intersubunit signal transduction, essential for ClpB chaperone activities.
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Dynamic structural states of ClpB involved in its disaggregation function. 査読あり
Takayuki Uchihashi, Yo-hei Watanabe, Yosuke Nakazaki, Takashi Yamasaki, Hiroki Watanabe, Takahiro Maruno, Kentaro Ishii, Susumu Uchiyama, Chihong Song, Kazuyoshi Murata, Ryota Iino, Toshio Ando
Nature communications 9 2147 2018年6月
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担当区分：筆頭著者, 責任著者
The ATP-dependent bacterial protein disaggregation machine, ClpB belonging to the AAA+ superfamily, refolds toxic protein aggregates into the native state in cooperation with the cognate Hsp70 partner. The ring-shaped hexamers of ClpB unfold and thread its protein substrate through the central pore. However, their function-related structural dynamics has remained elusive. Here we directly visualize ClpB using high-speed atomic force microscopy (HS-AFM) to gain a mechanistic insight into its disaggregation function. The HS-AFM movies demonstrate massive conformational changes of the hexameric ring during ATP hydrolysis, from a round ring to a spiral and even to a pair of twisted half-spirals. HS-AFM observations of Walker-motif mutants unveil crucial roles of ATP binding and hydrolysis in the oligomer formation and structural dynamics. Furthermore, repressed and hyperactive mutations result in significantly different oligomeric forms. These results provide a comprehensive view for the ATP-driven oligomeric-state transitions that enable ClpB to disentangle protein aggregates.
DOI： 10.1038/s41467-018-04587-w
その他リンク： http://www.nature.com/ncomms/
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Fusion protein analysis reveals the precise regulation between Hsp70 and Hsp100 during protein disaggregation. 査読あり
Sayaka Hayashi, Yosuke Nakazaki, Kei Kagii, Hiromi Imamura, Yo-hei Watanabe
Scientific Reports 7 ( 1 ) 8648 2017年8月
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Analysis of the Cooperative ATPase Cycle of the AAA+ Chaperone ClpB from Thermus thermophilus by Using Ordered Heterohexamers with an Alternating Subunit Arrangement 査読あり
Yamasaki Takashi、Oohata Yukiko、Nakamura Toshiki、Watanabe Yo-hei
J Biol Chem. 290 ( 15 ) 9789 - 9800 2015年4月
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ClpB chaperone passively threads soluble denatured proteins through its central pore 査読あり
Nakazaki Yosuke, Watanabe Yo-hei
Genes to Cells 19 ( 12 ) 891 - 900 2014年12月
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Conformational transition of the lid helix covering the protease active site is essential for the ATP-dependent protease activity of FtsH 査読あり
Suno R, Shimoyama M, Abe A, Shimamura T, Shimodate N, Watanabe YH, Akiyama Y, Yoshida M
FEBS Lett. 586 ( 19 ) 3117 - 3121 2012年9月
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Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein. 査読あり
Mizuno S, Nakazaki Y, Yoshida M, Watanabe YH.
FEBS J. 279 ( 8 ) 1474 - 1484 2012年4月
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Roles of conserved arginines in ATP-binding domains of AAA+ chaperone ClpB from Thermus thermophilus. 査読あり
Yamasaki T, Nakazaki Y, Yoshida M, Watanabe YH.
FEBS J. 278 ( 13 ) 2395 - 2403 2011年7月
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担当区分：筆頭著者
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Temperature-dependent regulation of Thermus thermophilus DnaK/DnaJ chaperones by DafA protein. 査読あり
Mizutani T, Nemoto S, Yoshida M, Watanabe YH.
Genes Cells 14 ( 12 ) 1405 - 1413 2009年12月
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担当区分：筆頭著者
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Stability of the two wings of the coiled-coil domain of ClpB chaperone is critical for its disaggregation activity. 査読あり
Watanabe YH, Nakazaki Y, Suno R, Yoshida M.
Biochem. J. 421 ( 1 ) 71 - 77 2009年6月
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担当区分：筆頭著者
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ATP binding to nucleotide binding domain (NBD)1 of the ClpB chaperone induces motion of the long coiled-coil, stabilizes the hexamer, and activates NBD2. 査読あり
Watanabe YH, Takano M, Yoshida M.
J. Biol. Chem. 280 ( 26 ) 24562 - 24567 2005年7月
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Trigonal DnaK-DnaJ complex versus free DnaK and DnaJ: heat stress converts the former to the latter, and only the latter can do disaggregation in cooperation with ClpB. 査読あり
Watanabe YH, Yoshida M.
J. Biol. Chem. 279 ( 16 ) 15723 - 15727 2004年4月
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The structure of ClpB: a molecular chaperone that rescues proteins from an aggregated state. 査読あり 国際共著
Lee S, Sowa ME, Watanabe YH, Sigler PB, Chiu W, Yoshida M, Tsai FT.
Cell 115 ( 2 ) 229 - 240 2003年10月
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Roles of the two ATP binding sites of ClpB from Thermus thermophilus. 査読あり
Watanabe YH, Motohashi K, Yoshida M.
J. Biol. Chem. 277 ( 8 ) 5804 - 5809 2002年2月
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Dual nature of the distal histidine residue in the autoxidation reaction of myoglobin and hemoglobin comparison of the H64 mutants. 査読あり
Suzuki T, Watanabe YH, Nagasawa M, Matsuoka A, Shikama K.
Eur. J. Biochem. 267 ( 20 ) 6166 - 6174 2000年10月
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Heat-inactivated proteins managed by DnaKJ-GrpE-ClpB chaperones are released as a chaperonin-recognizable non-native form. 査読あり
Watanabe YH, Motohashi K, Taguchi H, Yoshida M.
J. Biol. Chem. 275 ( 17 ) 12388 - 12392 2000年4月
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Heat-inactivated proteins are rescued by the DnaK.J-GrpE set and ClpB chaperones. 査読あり
Motohashi K, Watanabe Y, Yohda M, Yoshida M.
Proc. Natl. Acad. Sci. U. S. A. 96 ( 13 ) 7184 - 7189 1999年6月
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African elephant myoglobin with an unusual autoxidation behavior: comparison with the H64Q mutant of sperm whale myoglobin. 査読あり
Tada T, Watanabe YH, Matsuoka A, Ikeda-Saito M, Imai K, Ni-hei Y, Shikama K.
Biochim. Biophys. Acta. 1387 165 - 176 1998年9月
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