2012 Fiscal Year Final Research Report
Development of the substrate identification method against budding yeast SCF complex using site-specific photocross-linking in vivo
| Project/Area Number |
23657088
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Functional biochemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
KAMURA Takumi 名古屋大学, 理学研究科, 教授 (40333455)
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| Project Period (FY) |
2011 – 2012
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| Keywords | タンパク質分解 |
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| Research Abstract |
Ubiquitylation and subsequent proteasomal degradation of regulatory proteins control a variety of cellular processes. The E3s are responsible for recognizing and recruiting target proteins for polyubiquitylation. Relationship between E3s and substrates are largely uncharacterized because of the technical difficulty. In this study, we perform site-specific photocross-linking in vivo to identify specific substrates of SCF complex. Hard3 forms complex with Hrd1 and functions as a E3. We identify the specific binding region of Hrd3 with Hrd1 using site-specific photocross-linking in vivo. However, in these processes, we recognize that this method is not suitable for the detection of novel binding protein, because of the lacking of the binding region information. Then we perform yeast two-hybrid screening and identify p50 as a candidate of SCFmet30 substrate. Stability of p50 is regulated by SCFmet30 in the cell cycle dependent manner. We are currently investigating the functional meaning of this degradation.
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[Journal Article] Activation of double-stranded RNA-activated protein kinase (PKR) by interferon-stimulated gene 15 (ISG15) modification down-regulates protein translation2013
Author(s)
Okumura, F., Okumura, AJ., Uematsu, K., Hatakeyama, S., Zhang, DE., Kamura, T
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Journal Title
J Biol Chem
Volume: 288(4)
Pages: 2839-47
DOI
Peer Reviewed
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[Journal Article] Non-SCF type F-box protein Roy1/Ymr258c interacts with a Rab5-like GTPase Ypt52 and inhibits Ypt52 function2011
Author(s)
Liu, Y., Nakatsukasa, K., Kotera, M., Kanada, A., Nishimura, T., Kishi, T., Mimura, S., Kamura, T
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Journal Title
Mol. Biol. Cell
Volume: 22
Pages: 1575-1584
Peer Reviewed
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