| Project/Area Number |
12480192
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
KITAGAWA Masatoshi Hamamatsu University School of Medicine Professor, 医学部, 教授 (50294971)
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| Co-Investigator(Kenkyū-buntansha) |
KITAGAWA Kyoko Hamamatsu University School of Medicine, Assistant Professor, 医学部, 助手 (20299605)
UCHIDA Chiharu Hamamatsu University School of Medicine, Assistant Professor, 医学部, 助手 (60223567)
ODA Toshiaki Hamamatsu University School of Medicine, Associate Professor, 医学部, 助教授 (90126805)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2001: ¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 2000: ¥7,900,000 (Direct Cost: ¥7,900,000)
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| Keywords | p27^<Kip1> / Ubiquitin ligase / Gene therapy / prognosis / RB protein / Cell cycle / p27^<kip1> / ユビキチン-プロテアソーム / リン酸化 |
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| Research Abstract |
In this study, we analyzed the molecular mechanisms of proteolytic degradation of two major tumor suppressor gene products such as p27^<Kip1> and RB protein. Fast of all, we tried to identify the ubiquitin ligase for RB protein and analyze its function in malignant transformation process (2). The other project is identification of the down-regulation mechanisms of p27^<Kip1> (1).
(1) Down-regulation mechanisms of p27^<Kip1>
We as well as the other groups reported that p27^<Kip1> is efficiently degraded in human tumors with poor prognosis. Target disruption of Skp2, a ubiquitin ligase for p27^<Kip1>, resulted to decrease degradation of p27^<Kip1> in vivo. However, another mechanism of p27Kip1 degradation was stronglt suggested. We have reported that are down regulated by two mechanisms, one is ubiquitin dependent pathway and the other is site specific cleavage. We should identify the cleavage enzyme. In this study, we identified Ser10 as a major phosphorylation site for p27^<Kip1>. The phosphorylation at Ser10 stabilized p27^<Kip1> in cultured cells.
(2) Identification of ubiquitin ligase for RB protein
We established in vitro ubiquitination assay of RB protein and successfully obtain the candidate protein for ubiquitin ligase for RB protein using biochemical approach. In this study, it was confirmed by in vivo ubiquitination analysis and pulse chase experiment. It may be a novel mechanisms in malignant transformation via proteolytic degradation of RB protein.
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