Updated on 2023/03/17

写真a

 
KAMURA, Takumi
 
Organization
Graduate School of Science Professor
Graduate School
Graduate School of Science
Undergraduate School
School of Science
Title
Professor
Contact information
メールアドレス

Degree 1

  1. 医学博士 ( 1996.9   九州大学 ) 

Research Interests 1

  1. protein degradation

Research Areas 1

  1. Others / Others  / Molecular Biology

Current Research Project and SDGs 1

  1. Clarification of cellular processes regulated by ubiquitin-proteasome system

Research History 1

  1. Nagoya University   Graduate School of Science   Professor

    2005.11

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    Country:Japan

Education 1

  1. Kyushu University   Faculty of Medicine

    1982.4 - 1988.3

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    Country: Japan

Professional Memberships 1

  1. The Molecular Biology Society of Japan

 

Papers 45

  1. Proteolysis of adaptor protein Mmr1 during budding is necessary for mitochondrial homeostasis in Saccharomyces cerevisiae Reviewed

    Obara Keisuke, Yoshikawa Taku, Yamaguchi Ryu, Kuwata Keiko, Nakatsukasa Kunio, Nishimura Kohei, Kamura Takumi

    NATURE COMMUNICATIONS   Vol. 13 ( 1 )   2022.4

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/s41467-022-29704-8

    Web of Science

  2. Regulator of Awn Elongation 3, an E3 ubiquitin ligase, is responsible for loss of awns during African rice domestication Reviewed International coauthorship

    Kanako Bessho-Uehara, Kengo Masuda, Diane R Wang, Rosalyn B Angeles-Shim, Keisuke Obara, Keisuke Nagai, Riri Murase, Shin-Ichiro Aoki, Tomoyuki Furuta, Kotaro Miura, Jianzhong Wu, Yoshiyuki Yamagata, Hideshi Yasui, Michael B Kantar, Atsushi Yoshimura, Takumi Kamura, Susan R McCouch, Motoyuki Ashikari

    Proc Natl Acad Sci U S A   Vol. 120 ( 4 ) page: e2207105120   2023.1

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1073/pnas.2207105120

  3. A tonoplast-localized magnesium transporter is crucial for stomatal opening in Arabidopsis under high Mg2+ conditions

    Inoue Shin-ichiro, Hayashi Maki, Huang Sheng, Yokosho Kengo, Gotoh Eiji, Ikematsu Shuka, Okumura Masaki, Suzuki Takamasa, Kamura Takumi, Kinoshita Toshinori, Ma Jian Feng

    NEW PHYTOLOGIST   Vol. 236 ( 3 ) page: 864 - 877   2022.11

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    Language:Japanese  

    DOI: 10.1111/nph.18410

    Web of Science

  4. Triacylglycerol lipase Tgl4 is a stable protein and its dephosphorylation is regulated in a cell cycle-dependent manner in Saccharomyces cerevisiae Reviewed

    Nakatsukasa Kunio, Fujisawa Munetaka, Yang Xiaotan, Kawarasaki Tomoyuki, Okumura Fumihiko, Kamura Takumi

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   Vol. 626   page: 85 - 91   2022.10

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    DOI: 10.1016/j.bbrc.2022.08.022

    Web of Science

  5. Breaking the clip for cargo unloading from motor proteins: mechanism and significance

    Obara Keisuke, Kamura Takumi

    MICROBIAL CELL   Vol. 9 ( 6 ) page: 133 - 135   2022.6

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    DOI: 10.15698/mic2022.06.779

    Web of Science

  6. A positive genetic selection for transmembrane domain mutations in HRD1 underscores the importance of Hrd1 complex integrity during ERAD

    Nakatsukasa Kunio, Wigge Sylvia, Takano Yuki, Kawarasaki Tomoyuki, Kamura Takumi, Brodsky Jeffrey L.

    CURRENT GENETICS   Vol. 68 ( 2 ) page: 227 - 242   2022.4

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  7. Dot6/Tod6 degradation fine-tunes the repression of ribosome biogenesis under nutrient-limited conditions

    Kusama Kino, Suzuki Yuta, Kurita Ena, Kawarasaki Tomoyuki, Obara Keisuke, Okumura Fumihiko, Kamura Takumi, Nakatsukasa Kunio

    ISCIENCE   Vol. 25 ( 3 )   2022.3

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    Authorship:Corresponding author   Language:English  

    DOI: 10.1016/j.isci.2022.103986

    Web of Science

  8. ZSWIM8 is a myogenic protein that partly prevents C2C12 differentiation Reviewed International coauthorship

    Okumura Fumihiko, Oki Nodoka, Fujiki Yuha, Ikuta Rio, Osaki Kana, Hamada Shun, Nakatsukasa Kunio, Hisamoto Naoki, Hara Taichi, Kamura Takumi

    SCIENTIFIC REPORTS   Vol. 11 ( 1 )   2021.10

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/s41598-021-00306-6

    Web of Science

  9. The Rim101 pathway mediates adaptation to external alkalization and altered lipid asymmetry: hypothesis describing the detection of distinct stresses by the Rim21 sensor protein

    Obara Keisuke, Kamura Takumi

    CURRENT GENETICS   Vol. 67 ( 2 ) page: 213 - 218   2021.4

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    DOI: 10.1007/s00294-020-01129-0

    Web of Science

  10. The Rim101 pathway mediates adaptation to external alkalization and altered lipid asymmetry: hypothesis describing the detection of distinct stresses by the Rim21 sensor protein (vol

    Obara Keisuke, Kamura Takumi

    CURRENT GENETICS   Vol. 67 ( 2 ) page: 219 - 219   2021.4

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  11. A super-sensitive auxin-inducible degron system with an engineered auxin-TIR1 pair

    Nishimura Kohei, Yamada Ryotaro, Hagihara Shinya, Iwasaki Rie, Uchida Naoyuki, Kamura Takumi, Takahashi Koji, Torii Keiko U., Fukagawa Tatsuo

    NUCLEIC ACIDS RESEARCH   Vol. 48 ( 18 )   2020.10

  12. Rapid turnover of transcription factor Rim101 confirms a flexible adaptation mechanism against environmental stress inSaccharomyces cerevisiae Reviewed

    Obara Keisuke, Higuchi Mai, Ogura Yuki, Nishimura Kohei, Kamura Takumi

    GENES TO CELLS   Vol. 25 ( 10 ) page: 651 - 662   2020.10

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    DOI: 10.1111/gtc.12801

    Web of Science

  13. Cul5-type Ubiquitin Ligase KLHDC1 Contributes to the Elimination of Truncated SELENOS Produced by Failed UGA/Sec Decoding Reviewed

    Okumura Fumihiko, Fujiki Yuha, Oki Nodoka, Osaki Kana, Nishikimi Akihiko, Fukui Yoshinori, Nakatsukasa Kunio, Kamura Takumi

    ISCIENCE   Vol. 23 ( 3 )   2020.3

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    DOI: 10.1016/j.isci.2020.100970

    Web of Science

  14. N-glycosylation of Rim21 at an Unconventional Site Fine-tunes Its Behavior in the Plasma Membrane Reviewed

    Obara Keisuke, Kotani Tetsuya, Nakatogawa Hitoshi, Kihara Akio, Kamura Takumi

    CELL STRUCTURE AND FUNCTION   Vol. 45 ( 1 ) page: 1 - 8   2020

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    DOI: 10.1247/csf.19021

    Web of Science

  15. Hepatic Sdf2l1 controls feeding-induced ER stress and regulates metabolism

    Sasako Takayoshi, Ohsugi Mitsuru, Kubota Naoto, Itoh Shinsuke, Okazaki Yukiko, Terai Ai, Kubota Tetsuya, Yamashita Satoshi, Nakatsukasa Kunio, Kamura Takumi, Iwayama Kaito, Tokuyama Kumpei, Kiyonari Hiroshi, Furuta Yasuhide, Shibahara Junji, Fukayama Masashi, Enooku Kenichiro, Okushin Kazuya, Tsutsumi Takeya, Tateishi Ryosuke, Tobe Kazuyuki, Asahara Hiroshi, Koike Kazuhiko, Kadowaki Takashi, Ueki Kohjiro

    NATURE COMMUNICATIONS   Vol. 10   2019.2

  16. Insulin-like growth factor 1 receptor stabilizes the ETV6-NTRK3 chimeric oncoprotein by blocking its KPC1/Rnf123-mediated proteasomal degradation Reviewed International coauthorship

    Cristina E Tognon, Bo Rafn, Naniye Malli Cetinbas, Takumi Kamura, Genny Trigo, Barak Rotblat, Fumihiko Okumura, Masaki Matsumoto, Christine Chow, Monika Davare, Michael Pollak, Thibault Mayor, Poul H Sorensen

    J Biol Chem   Vol. 293 ( 32 ) page: 12502 - 12515   2018.8

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1074/jbc.RA117.000321

  17. The HECT-type ubiquitin ligase Tom1 contributes to the turnover of Spo12, a component of the FEAR network, in G2/M phase Reviewed

    Nakatsukasa Kunio, Sone Megumi, Alemayehu Dawit Hailu, Okumura Fumihiko, Kamura Takumi

    FEBS LETTERS   Vol. 592 ( 10 ) page: 1716-1724   2018.5

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    DOI: 10.1002/1873-3468.13066

    Web of Science

  18. Ubiquitin ligase SPSB4 diminishes cell repulsive responses mediated by EphB2 Reviewed

    Okumura Fumihiko, Joo-Okumura Akiko, Obara Keisuke, Petersen Alexander, Nishikimi Akihiko, Fukui Yoshinori, Nakatsukasa Kunio, Kamura Takumi

    MOLECULAR BIOLOGY OF THE CELL   Vol. 28 ( 24 ) page: 3532-3541   2017.11

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    DOI: 10.1091/mbc.E17-07-0450

    Web of Science

  19. Hypoxia-inducible factor-2a stabilizes the von Hippel-Lindau (VHL) disease suppressor, Myb-related protein 2 Reviewed

    Okumura Fumihiko, Joo-Okumura Akiko, Nakatsukasa Kunio, Kamura Takumi

    PLOS ONE   Vol. 12 ( 4 )   2017.4

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    DOI: 10.1371/journal.pone.0175593

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  20. Subcellular Fractionation Analysis of the Extraction of Ubiquitinated Polytopic Membrane Substrate during ER-Associated Degradation. Reviewed

    Nakatsukasa, K., Kamura, T.

    PloS One   Vol. 11 ( 2 ) page: e0148327   2016.2

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    DOI: 10.1371/journal.pone.0148327

  21. The role of cullin 5-containing ubiquitin ligases. Reviewed

    Okumura, F., Okumura, AJ., Nakatsukasa, K., Kamura, T

    Cell Division     2016

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    DOI: 10.1186/s13008-016-0016-3

  22. ASB7 regulates spindle dynamics and genome integrity by targeting DDA3 for proteasomal degradation. Reviewed

    Uematsu, K., Okumura, F., Tonogai, S., Okumura, AJ., Alemayehu, DH., Nishikimi, A., Fukui, Y., Nakatsukasa, K., and Kamura, T

      Vol. 215 ( 1 ) page: 95-106   2016

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    DOI: 10.1083/jcb.201603062.

  23. Parallel regulation of VHL disease by pVHL-mediated degradation of B-Myb and HIF-α. Reviewed

    Okumura, F., Uematsu, K., Byrne, SD., Hirano, M., Joo-Okumura, A., Nishikimi, A., Shuin. T., Fukui, Y., Nakatsukasa, K., Kamura, T

    Mol.Cell. Biol.   Vol. 36 ( 12 ) page: 1803-17   2016

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    DOI: 10.1128/MCB.00067-16

  24. The ubiquitin ligase SCFUcc1 acts as a metabolic switch for the glyoxylate cycle Reviewed

    1. Nakatsukasa, K., Nishimura, T., Byrne, D., Okamoto, M., Takahashi-Nakaguchi, A., Chibana, H., Okumura, F. and Kamura, T

    Molecular Cell   Vol. 59 ( 1 ) page: 22-34   2015.7

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    Despite the crucial role played by the glyoxylate cycle in the virulence of pathogens, seed germination in plants, and sexual development in fungi, we still have much to learn about its regulation. Here, we show that a previously uncharacterized SCF(Ucc1) ubiquitin ligase mediates proteasomal degradation of citrate synthase in the glyoxylate cycle to maintain metabolic homeostasis in glucose-grown cells. Conversely, transcription of the F box subunit Ucc1 is downregulated in C2-compound-grown cells, which require increased metabolic flux for gluconeogenesis. Moreover, in vitro analysis demonstrates that oxaloacetate regenerated through the glyoxylate cycle induces a conformational change in citrate synthase and inhibits its recognition and ubiquitination by SCF(Ucc1), suggesting the existence of an oxaloacetate-dependent positive feedback loop that stabilizes citrate synthase. We propose that SCF(Ucc1)-mediated regulation of citrate synthase acts as a metabolic switch for the glyoxylate cycle in response to changes in carbon source, thereby ensuring metabolic versatility and flexibility.

    DOI: 10.1016/j.molcel.2015.04.013

  25. The nutrient stress-induced small GTPase Rab5 contributes to the activation of vesicle trafficking and vacuolar activity. Reviewed

    3. Nakatsukasa, K., Kanada, A., Matsuzaki, M., Byrne, SD., Okumura, F., Kamura, T

    Journal of biological chemistry   Vol. 289 ( 30 ) page: 20970-20978   2014

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    DOI: 10.1074/jbc.M114.548297

  26. Integrated molecular analysis of clear-cell renal cell carcinoma Reviewed

    2. Sato Y1, Yoshizato T, Shiraishi Y, Maekawa S, Okuno Y, Kamura T, Shimamura T, Sato-Otsubo A, Nagae G, Suzuki H, Nagata Y, Yoshida K, Kon A, Suzuki Y, Chiba K, Tanaka H, Niida A, Fujimoto A, Tsunoda T, Morikawa T, Maeda D, Kume H, Sugano S, Fukayama M, Aburatani H, Sanada M, Miyano S, Homma Y, Ogawa S.

    Nature Genetics   Vol. 45 ( 8 ) page: 860-867   2013.8

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    DOI: 10.1038/ng.2699

  27. A stalled retrotranslocation complex reveals physical linkage between substrate recognition and proteasomal degradation during ER-associated degradation. Reviewed

    Nakatsukasa K, Brodsky JL, Kamura T.

    Molecular biology of the cell   Vol. 24 ( 11 ) page: 1765-1775   2013.6

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    DOI: 10.1091/mbc.E12-12-0907

  28. Activation of Double-stranded RNA-activated Protein Kinase (PKR) by Interferon-stimulated Gene 15 (ISG15) Modification Down-regulates Protein Translation. Reviewed

    Okumura F, Okumura AJ, Uematsu K, Hatakeyama S, Zhang DE, Kamura T

    Journal of biological chemistry   Vol. 288 ( 4 ) page: 2839-2847   2013.1

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    DOI: 10.1074/jbc.M112.401851

  29. Non-SCF type F-box protein Roy1/Ymr258c interacts with a Rab5-like GTPase Ypt52 and inhibits Ypt52 function. Reviewed

    1. Liu, Y., Nakatsukasa, K., Kotera, M., Kanada, A., Nishimura, T., Kishi, T., Mimura, S., Kamura, T.

    Mol. Biol. Cell   Vol. 22 ( 9 ) page: 1574-1584   2011.5

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    Skp1/Cul1/F-box (SCF)-type F-box proteins are a component of the Cullin-RING SCF ubiquitin E3 ligase, which is involved in numerous cellular processes. However, the function of non-SCF-type F-box proteins remains largely unknown. The Rab5-like small guanosine 5'-triphosphatase Vps21/Ypt51 is a key regulator of intracellular transportation; however, deletion of its isoforms, Ypt52 and Ypt53, results in only a modest inhibition of intracellular trafficking. The function of these proteins therefore remains largely elusive. Here we analyze the role of a previously uncharacterized non-SCF-type F-box protein, Roy1/Ymr258c, in cell growth and intracellular transport in Saccharomyces cerevisiae. Roy1 binds to Ypt52 under physiological conditions, and Skp1 is indispensable for the association of Roy1 with Ypt52. The vps21Δ yeast cells exhibit severe deficiencies in cell growth and intracellular trafficking, whereas simultaneous deletion of roy1 alleviates the defects caused by deletion of vps21. However, additional disruption of ypt52 in roy1Δvps21Δ cells largely suppresses the cell growth and trafficking observed in roy1Δvps21Δ cells. We demonstrate that Roy1 interacts with guanosine 5'-diphosphate-bound and nucleotide-free Ypt52 and thereby inhibits the formation of guanosine 5'-triphosphate-bound, active Ypt52. These results thus indicate that Roy1 negatively modulates cell viability and intracellular transport by suppressing Ypt52.

  30. Cul8/Rtt101 forms a variety of protein complexes that regulate DNA damage response and transcriptional silencing Reviewed

    1. Mimura, S., Yamaguchi, T., Ishii, S., Noro, E., Katsura, T., Obuse, C., Kamura, T

    Journal of biological chemistry   Vol. 285 ( 13 ) page: 9858-9867   2010.3

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    The budding yeast, Saccharomyces cerevisiae, has three cullin proteins, which act as platforms for Cullin-based E3 ubiquitin ligases. Genetic evidence indicates that Cul8, together with Mms1, Mms22, and Esc4, is involved in the repair of DNA damage that can occur during DNA replication. Cul8 is thought to form a complex with these proteins, but the composition and the function of Cul8-based E3 ubiquitin ligases remain largely uncharacterized. Herein, we report a comprehensive biochemical analysis of Cul8 complexes. Cul8 was found to form a Cul8-Mms1-Mms22-Esc4 complex under physiological conditions, with Mms1 bridging Cul8 and Mms22 and Mms22 bridging Mms1 and Esc4. Domain analysis demonstrated that the N-terminal region of Mms1 and the C-terminal region of Mms22 are required for the Mms1-Mms22 interaction, whereas the N-terminal region of Mms22 is required for the Mms22-Esc4 interaction. We also found other Cul8-Mms1-binding proteins Ctf4, Esc2, and Orc5 using yeast two-hybrid screening. Esc4 and Ctf4 bound to Mms22 directly and bound to Cul8-Mms1 in the presence of Mms22, whereas Esc2 and Orc5 interacted with both Cul8 and Mms1, independently. We found that Cul8, Mms1, and Mms22 participated in the regulation of transcriptional silencing of yeast telomeres. These results suggest that Cul8-Mms1, as part of various protein complexes, is involved in the regulation of chromatin metabolism.

  31. *SCFDia2 regulates DNA replication forks during S-phase in budding yeast. Reviewed

    Mimura, S., Komata, M., Kishi, T., Shirahige, K., Kamura, T.

    EMBO J.   Vol. 28 ( 23 ) page: 3693-3705   2009.12

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    Dia2 is an F-box protein, which is involved in the regulation of DNA replication in the budding yeast Saccharomyces cerevisiae. The function of Dia2, however, remains largely unknown. In this study, we report that Dia2 is associated with the replication fork and regulates replication fork progression. Using modified yeast two-hybrid screening, we have identified components of the replisome (Mrc1, Ctf4 and Mcm2), as Dia2-binding proteins. Mrc1 and Ctf4 were ubiquitinated by SCF(Dia2) both in vivo and in vitro. Domain analysis of Dia2 revealed that the leucine-rich repeat motif was indispensable for the regulation of replisome progression, whereas the tetratricopeptide repeat (TPR) motif was involved in the interaction with replisome components. In addition, the TPR motif was shown to be involved in Dia2 stability; deleting the TPR stabilized Dia2, mimicking the effect of DNA damage. ChIP-on-chip analysis illustrated that Dia2 localizes to the replication fork and regulates fork progression on hydroxyurea treatment. These results demonstrate that Dia2 is involved in the regulation of replisome activity through a direct interaction with replisome components.

  32. *A longevity protein, Lag2, interacts with SCF complex and regulates SCF function. Reviewed

    Liu, Y., Mimura, S., Kishi, T., Kamura, T.

    EMBO J.   Vol. 28 ( 21 ) page: 3366-3377   2009.11

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    SCF-type E3-ubiquitin ligases control numerous cellular processes through the ubiquitin-proteasome pathway. However, the regulation of SCF function remains largely uncharacterized. Here, we report a novel SCF complex-interacting protein, Lag2, in Saccharomyces cerevisiae. Lag2 interacts with the SCF complex under physiological conditions. Lag2 negatively controls the ubiquitylation activities of SCF E3 ligase by interrupting the association of Cdc34 to SCF complex. Overexpression of Lag2 increases unrubylated Cdc53, whereas deletion of lag2, together with the deletions of dcn1 and jab1, results in the accumulation of Rub1-modified Cdc53. In vitro rubylation assays show that Lag2 inhibits the conjugation of Rub1 to Cdc53 in competition with Dcn1, which suggest that Lag2 down-regulates the rubylation of Cdc53 rather than promoting derubylation. Furthermore, Dcn1 hinders the association of Lag2 to Cdc53 in vivo. Finally, the deletion of lag2 combined with the deletion of either dcn1 or rub1 suppresses the growth of yeast cells. These observations thus indicate that Lag2 has a significant function in regulating the SCF complex by controlling its ubiquitin ligase activities and its rubylation cycle.

  33. *Degradation of phosphorylated p53 by viral protein-ECS E3 ligase complex. Reviewed

    3. Sato, Y., Kamura, T., Shirata, N., Murata, T., Kudoh, A., Iwahori, S., Nakayama, S., Isomura, H., Nishiyama, Y., Tsurumi T.

    PLoS Pathog   Vol. 5 ( 7 ) page: e1000530   2009.7

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    p53-signaling is modulated by viruses to establish a host cellular environment advantageous for their propagation. The Epstein-Barr virus (EBV) lytic program induces phosphorylation of p53, which prevents interaction with MDM2. Here, we show that induction of EBV lytic program leads to degradation of p53 via an ubiquitin-proteasome pathway independent of MDM2. The BZLF1 protein directly functions as an adaptor component of the ECS (Elongin B/C-Cul2/5-SOCS-box protein) ubiquitin ligase complex targeting p53 for degradation. Intringuingly, C-terminal phosphorylation of p53 resulting from activated DNA damage response by viral lytic replication enhances its binding to BZLF1 protein. Purified BZLF1 protein-associated ECS could be shown to catalyze ubiquitination of phospho-mimetic p53 more efficiently than the wild-type in vitro. The compensation of p53 at middle and late stages of the lytic infection inhibits viral DNA replication and production during lytic infection, suggesting that the degradation of p53 is required for efficient viral propagation. Taken together, these findings demonstrate a role for the BZLF1 protein-associated ECS ligase complex in regulation of p53 phosphorylated by activated DNA damage signaling during viral lytic infection.

  34. *Mammalian Elongin A complex mediates DNA-damage-induced ubiquitylation and degradation of Rpb1. Reviewed

    Yasukawa T, Kamura T, Kitajima S, Conaway RC, Conaway JW, Aso T.

    EMBO J.   Vol. 27 ( 24 ) page: 3256-3266   2008.12

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    The Elongin complex stimulates the rate of transcription elongation by RNA polymerase II (pol II) by suppressing transient pausing of the pol II at many sites along the DNA. Elongin is composed of a transcriptionally active A subunit and two small regulatory B and C subunits, which can form an isolable Elongin BC subcomplex. Here, we have shown that both the ubiquitylation and proteasomal degradation of the largest subunit of pol II (Rpb1) following UV-irradiation are significantly suppressed in Elongin A-deficient cells; however, in both cases suppression is rescued by transfection of wild-type Elongin A. Moreover, we have demonstrated that the Elongin A-Elongin BC complex is capable of assembling with the Cul5/Rbx2 module, and that this hetero-pentamer complex efficiently ubiquitylates Rpb1 in vitro. Mechanistic studies indicate that colocalization of Elongin A and Cul5 in cells and the interaction of Elongin A with the Ser5-phosphorylated form of Rpb1 are strongly enhanced following UV-irradiation. Taken together, our results suggest that mammalian Elongin A is directly involved in ubiquitylation and degradation of Rpb1 following DNA damage.

  35. Fbxw7 contributes to tumor suppression by targeting multiple proteins for ubiquitin-dependent degradation. Reviewed

    Fujii, Y., Yada, M., Nishiyama, M., Kamura, T., Takahashi, H., Tsunematsu, R., Susaki, E., Nakagawa, T., Matsumoto, A., Nakayama, KI.

    Cancer Science   Vol. 97 ( 8 ) page: 729-736   2006.8

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  36. *Fbxw8 is essential for Cul1-Cul7 complex formation and for placental development. Reviewed

    Tsunematsu, R., Nishiyama, M., Kotoshiba, S., Saiga, T., Kamura, T., Nakayama, KI.

    Molecular and Cellular Biology   Vol. 26 ( 16 ) page: 6157-6169   2006.8

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  37. Role of the UBL-UBA protein KPC2 in degradation of p27 at G1 phase of the cell cycle. Reviewed

    Hara, T., Kamura, T., Kotoshiba, S., Takahashi, H., Fujiwara, K., Onoyama, I., Shirakawa, M., Mizushima, N., Nakayama, KI.

    Molecular and Cellular Biology   Vol. 25 ( 21 ) page: 9292-9303   2005.11

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  38. Molecular dissection of the interaction between p27 and KPC, the ubiquitin ligase that regulates proteolysis of p27 in G1 phase. Reviewed

    Kotoshiba, S., Kamura, T., Hara, T., Ishida, N., Nakayama, KI.

    Journal of Biological Chemistry   Vol. 280 ( 18 ) page: 17694-17700   2005.5

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  39. Functional regulation of FEZ1 by the U-box-type ubiquitin ligase E4B contributes to neuritogenesis. Reviewed

    Okumura, F., Hatakeyama, S., Matsumoto, M., Kamura, T., Nakayama, K.I.

    Journal of Biological Chemistry   Vol. 279 ( 51 ) page: 53533-53543   2004.12

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  40. VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases. Reviewed

    Kamura, T., Maenaka, K., Kotoshiba, S., Matsumoto, M., Kohda, D., Conaway, R.C., Conaway, J.W., Nakayama, K.I.

    Genes & Development   Vol. 18 ( 24 ) page: 3055-3065   2004.12

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  41. Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27Kip1 at G1 phase. Reviewed

    Kamura, T., Hara, T., Matsumoto, M., Ishida, N., Okumura, F., Hatakeyama, S., Yoshida, M., Nakayama, K., Nakayama, K.I.

    Nature Cell Biology   Vol. 6 ( 12 ) page: 1229-1235   2004.12

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  42. Evidence for impaired Skp2-dependent degradation of p27 in terminal differentiation. Reviewed

    Tamamori-Adachi, M., Hayashida, K., Nobori, K., Omizu, C., Yamada, K., Sakamoto, N., Kamura, T., Fukuda, K., Ogawa, S., Nakayama, K.I., Kitajima, S.

    Journal of Biological Chemistry   Vol. 279 ( 48 ) page: 50429-50436   2004.11

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  43. Identification of elongin C and Skp1 sequences that determine cullin selection. Reviewed

    Yan, Q., Kamura, T., Cai, Y., Jin, J., Ivan, M., Mushegian, A., Conaway, R.C., Conaway ,J.W.

    Journal of Biological Chemistry   Vol. 279 ( 41 ) page: 43019-43026   2004.10

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  44. U-box protein carboxyl terminus of Hsc70-interacting protein (CHIP) mediates poly-ubiquitylation preferentially on four-repeat Tau and is involved in neurodegeneration of tauopathy. Reviewed

      Vol. 91 ( 2 ) page: 299-307   2004.10

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  45. Phosphorylation- dependent degradation of c-Myc is mediated by the F-box protein Fbw7. Reviewed

    Yada, M., Hatakeyama, S., Kamura, T., Nishiyama, M., Tsunematsu, R., Imaki, H., Ishida, N., Okumura, F., Nakayama, K., Nakayama, K.I.

    EMBO Journal   Vol. 23 ( 10 ) page: 2116-2125   2004.5

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Presentations 5

  1. ユビキチンシステムと発癌・細胞周期

    嘉村 巧

    第68回日本血液学会総会・第48回日本臨床血液学会総会・合同総会 

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    Event date: 2006.10

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

  2. Identification of an ubiquitin ligase, KPC, that regulates proteolysis of p27Kip1 at the G0-G1 transition. International conference

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    Event date: 2006.3

    Language:English   Presentation type:Oral presentation (invited, special)  

    Country:Japan  

  3. Cullin/Rbx型E3によるタンパク質分解機構

    嘉村 巧、中山 敬一

    第28回日本分子生物学会年会 

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    Event date: 2005.12

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Country:Japan  

  4. VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases. International conference

    The second Cold Spring Harbor Laboratory meeting on The Ubiquitin Family. 

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    Event date: 2005.4

    Language:English   Presentation type:Oral presentation (general)  

  5. C-terminus of the conserved SOCS-box motif present in members of the SOCS-box protein families is indispensable for interaction with Cul5 and Rbx2. International conference

    6th International Symposium on von Hippel-Lindau Disease. 

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    Event date: 2004.5

    Language:English   Presentation type:Oral presentation (invited, special)  

    Country:Japan  

Research Project for Joint Research, Competitive Funding, etc. 4

  1. 複合体型ユビキチンリガーゼの機能解析

    2006

  2. 新規ユビキチン化酵素KPC複合体による細胞周期制御因子p27の分解機構の解析

    2006

  3. 異常タンパク質蓄積による神経変性疾患発症の分子機構の解明

    2006

  4. ユビキチンリガーゼの多様性の解析

    2006

KAKENHI (Grants-in-Aid for Scientific Research) 6

  1. Artificial cell cycle control using the AID method, a low-toxicity and rapid proteolytic system

    Grant number:20K21423  2020.7 - 2022.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Research (Exploratory)

    Kamura Takumi

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    Authorship:Principal investigator 

    Grant amount:\6500000 ( Direct Cost: \5000000 、 Indirect Cost:\1500000 )

    The AID method, a low-toxicity and rapid proteolytic system, was used to artificially control the cell cycle in cultured cells. Since cell cycle progression is positively regulated by CDK complexes and negatively regulated by CKI, the expression of these factors was controlled by the AID method and their effects on the cell cycle were examined. We found that in the Cdk1 AID cell line, the cell cycle is arrested in the G2 phase due to Cdk1 degradation. Similar results were obtained not only in the chicken DT40 cell line but also in mouse ES cells, suggesting that artificial cell cycle control can be achieved by regulating Cdk1 in a variety of cells.

  2. ユビキチン依存性タンパク質分解により制御される生命現象の解明

    Grant number:20H03208  2020.4 - 2023.3

    科学研究費助成事業  基盤研究(B)

    嘉村 巧

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    Grant amount:\17810000 ( Direct Cost: \13700000 、 Indirect Cost:\4110000 )

    ユビキチン・プロテアソーム系を介したタンパク質分解は、細胞周期進行など多岐にわたる生命現象に重要な働きを果たしている。現在までの精力的な研究により生体内に非常に多くのE3が存在し、基質特異性を決める役割を果たしていることが明らかになっている。その中でもSCF複合体が重要な役割を担っていることが予想されている。最近我々は出芽酵母の充実したリソースとデータベースを活用することにより、短寿命タンパク質から対応するE3を同定する方法を見出している。そこで本研究では、SCF複合体の新規機能解明を行ない、最終的にはユビキチン依存性タンパク質分解という観点から様々な生命現象を解明することを目的とする。
    ユビキチン・プロテアソーム系を介したタンパク質分解は、細胞周期進行やシグナル伝達など多岐にわたる生命現象に重要な働きを果たしている。現在までの精力的な研究により生体内に非常に多くのE3が存在し、基質特異性を決める役割を果たしていることが明らかになっている。その中でもSCF複合体(出芽酵母ではE3の約2割を占める)が重要な役割を担っていることが予想されているが、技術的困難さのため、酵素と基質の対応関係が明らかになっているのはごく僅かである。最近我々は出芽酵母の充実したリソースとデータベースを活用することにより、短寿命タンパク質(基質候補タンパク質)から対応するE3を同定する方法を見出している。この方法を用いて、すでにBdf2、Vhs1など複数のSCF複合体の新規基質の同定に成功している。そこで本研究では、この方法を用いてSCF複合体の新たな基質の同定・解析を進めること目的としている。われわれはストレス応答因子のひとつであるTmc1 (Trivalent Metalloid sensitive, Cuz1-relatedprotein)に注目し解析を進めている。Tmc1は半減期約40分と短寿命タンパク質であり、先行研究より、ストレス応答性転写因子Rpn4によって転写が活性化され、様々なストレス (カドミウム、DNA複製ストレスなど)応答時に発現量が制御されることが報告されている。出芽酵母E3欠失株、温度感受性変異株を用いた網羅的解析により、Tmc1がSCF複合体依存的に分解されることを見出した。さらにTmc1の機能解析を進め、①Tmc1がヒ素が存在する環境中で発現が増大すること、②TMC1欠損変異体は3価ヒ素に感受性を示すこと、③Tmc1はヒ素存在下においてARR2の転写を誘導することを見出した。現在、分解によるTmc1の機能制御機構を調べている最中である。
    われわれはSCF複合体の新規基質の同定を進めている。出芽酵母のデータベースより網羅的に短寿命タンパク質を選び、その分解を制御するE3の同定を進めている。その過程で、ストレス応答転写因子であるmc1がSCF複合体依存的に分解されることを見出し、その解析を進めているところであることより、研究はおおむね順調に進展していると判断した。
    1. 新規基質のF-boxタンパク質群による認識機構の解析 野生株およびE3欠失株より免疫沈降法で内在性の基質を精製する。質量分析でE3欠失株において増えている修飾およびその部位を同定する。野生型および修飾部位に変異を持つ基質の発現ベクターを作製し、酵母内でのF-boxタンパク質との結合を調べる。
    2. 新規基質に対する試験管内ユビキチン化反応の検討  野生型あるいは1.で同定した修飾部位に変異を持つ基質を大腸菌で発現させリコンビナントタンパク質を精製する。バキュロウイルス発現系を用いてSCF複合体を精製し、ユビキチン化反応に必要な酵素E1とE2そしてユビキチンさらにATPを加え基質と反応させ、試験管内で基質へのユビキチン化反応を起こすことができるかどうかを検討する。
    3. 新規基質に対する細胞内分解の検討  2.の試験管内ユビキチン化反応により得られた情報を細胞内で確認する。野生型あるいは変異型の基質とF-boxタンパク質を出芽酵母に発現させ、基質に対する抗体を用いて免疫沈降する。SDS-PAGEで展開した後、抗ユビキチン抗体でウェスタンブロットを行う。野生型の基質はユビキチン化されるのに対し、変異型の基質はユビキチン化されないはずである。また、シクロヘキシミドチェイス法により基質の半減期を測定する。
    4. 新規基質の分解制御による細胞生物学的影響の検討  F-boxタンパク質の過剰発現、遺伝子欠失によって基質の発現を制御することにより、どのような細胞生物学的変化が現れるかを解析する。
    5. データベースを用いた高等生物でのホモロジー検索および機能解析  上記研究で同定した基質を基にデータベース検索を行い高等生物にホモログが存在するかどうかを調べる。もしもホモログが存在すればそれらの機能解析を行う。

  3. 出芽酵母リン酸化酵素TDA1の糖シグナル伝達における役割の解明

    Grant number:19H04957  2019.4 - 2021.3

    科学研究費助成事業  新学術領域研究(研究領域提案型)

    嘉村 巧

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    Grant amount:\10530000 ( Direct Cost: \8100000 、 Indirect Cost:\2430000 )

    細胞内エネルギーの恒常性を維持することは正常な生命活動を行う上で重要である。その中心的な役割を担っているのは、出芽酵母においてはSnf1キナーゼでありエネルギー状態のセンサーとして機能し、糖シグナル伝達経路の活性を制御している。我々は、最近Tda1キナーゼも糖シグナル経路に関与していることを見出した。そこで本課題では、Tda1の糖シグナル伝達における役割に焦点を当てて研究を行う。
    細胞内エネルギーの恒常性を維持することは正常な生命活動を行う上で重要である。その中心的な役割を担っているのは、出芽酵母においてはSnf1キナーゼでありエネルギー状態のセンサーとして機能し、糖シグナル伝達経路の活性を制御している。われわれは、ユビキチンープロテアソーム依存性タンパク質分解により制御される生命現象の解明を目的の研究を進めているが、その過程で、出芽酵母キナーゼタンパク質Tda1の分解を制御するE3を新たに同定した。Tda1は、最近出芽酵母に3種類存在するヘキソキナーゼの1つであるHxk2をリン酸化することが示されているが、一方ではSnf1がHxk2をリン酸化するという報告もある。そこで本課題では、Tda1のE3による分解制御および糖シグナル伝達における役割に焦点を当てて研究を進めた。まずTda1がSCF複合体依存的に分解制御を受けていることを見出した。続いて低グルコースあるいは非発酵性炭素源環境下におけるSnf1によるTda1のリン酸化修飾を詳細に調べた。Snf1はSnf4およびGal83/Sip1/Sip2からなる複合体を形成しているが、この中でSnf1とSnf4はTda1のリン酸化に必要であったが、Gal83/Sip1/Sip2は不要であった。また試験管内リン酸化反応においてSnf1が直接Tda1をリン酸化修飾することを確認した。続いてHxk2のリン酸化修飾を検討した。その結果、低グルコース下で活性化したSnf1によりTda1がリン酸化され活性状態なることによりHxk2をリン酸化することを見出した。試験管内リン酸化反応においても同様な結果を得ている。これらのことよりTda1の発現量はユビキチン・プロテアソーム系によって調節されていること、そしてTda1はHxk2の活性を制御することにより糖シグナル伝達をコントロールしていることが明らかになった。
    令和2年度が最終年度であるため、記入しない。
    令和2年度が最終年度であるため、記入しない。

  4. Development of a lipid asymmetry biosensor that dramatically facilitates studies on biomembranes

    Grant number:18K19292  2018.6 - 2020.3

    Grant-in-Aid for Challenging Research (Exploratory)

    Kamura Takumi

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    Grant amount:\6240000 ( Direct Cost: \4800000 、 Indirect Cost:\1440000 )

    In this study, we tried to develop a biosensor that can report the state of lipid asymmetry in the plasma membrane. For this purpose, we focused on and utilized a lipid asymmetry sensor protein Rim21 which we previously identified in yeast. As a result, we developed a prototype of lipid asymmetry biosensor that can visualize alterations in lipid asymmetry in living yeast cells. In addition, we conducted a systematic mutagenesis approach of the prototype biosensor, and succeeded in developing a series of improved biosensors with higher S/N ratio.
    Using these biosensors, we monitored the state of lipid asymmetry in yeast cells exposed to environmental stresses. Interestingly, the state of lipid symmetry seemed to be altered under alkaline and salt stresses. Cells deleted for RIM21 were hypersensitive to alkaline and salt stresses, suggesting that these environmental stresses are sensed by Rim21 through alterations in the state of lipid asymmetry in the plasma membrane.

  5. Elucidation of novel function of Saccharomyces cerevisiae SCF complex

    Grant number:17H03652  2017.4 - 2020.3

    Kamura Takumi

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    Grant amount:\17420000 ( Direct Cost: \13400000 、 Indirect Cost:\4020000 )

    Proteolysis via the ubiquitin-proteasome system has attracted much attention as it has been revealed that it plays an important role in various life phenomena. E3, which determines substrate specificity, and SCF complexes in particular, are expected to have a variety of functions due to their large number. In the present study, we aimed to analyze a new function of the budding yeast SCF complex. We used genetic and biochemical methods to search for novel substrates of the SCF complex and identified the stress-responsive transcription factor Tmc1, which we found to be degraded in a SCF complex-dependent manner. We further found that Tmc1 adapts to arsenic stress by inducing the expression of Arr2.

  6. 細胞周期をコントロールするユビキチンリガーゼ群の機能解析

    2007

    科学研究費補助金  萌芽研究,課題番号:19657059

    嘉村 巧

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Teaching Experience (On-campus) 1

  1. Fundamentals of Biology II

    2011