| Project/Area Number |
16390072
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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 |
General medical chemistry
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| Research Institution | Nagahama Institute of Bio-Science and Technology (2005)
University of Tsukuba (2004) |
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Principal Investigator |
MIWA Masanao Nagahama Inst of Bio-Sci Tech, Bioscience, Professor, バイオサイエンス学部, 教授 (20012750)
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| Co-Investigator(Kenkyū-buntansha) |
KAMEMURA Kazuo Nagahama Inst Bio-Sci Tech, Bioscience, Lecturer, バイオサイエンス学部, 講師 (00399437)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2005: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 2004: ¥6,400,000 (Direct Cost: ¥6,400,000)
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| Keywords | Posttranslational modification / Poly(ADP-ribosyl)ation / Poly(ADP-ribose) glycohydrolase / Drosophila melanogaster / PARP / Centrosome / ポリADP-リボース / p53 / ポリADP-リボース分解酵素 / 神経変性 / 細胞周期 / 細胞分裂 |
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| Research Abstract |
We identified tow proteins that are poly(ADP-ribosyl)ated in vivo. They are p53 and nucleophosmin/B23. To identify the site of poly(ADP-ribosyl)ation of p53, we made the expression vector to produce GST-p53 fusion protein in E. coli. We confirmed the poly(ADP-ribosyl)ation of the entire product of p53 (aa1-aa393) by PARP-1. Currently we are preparing several GST-truncated p53 proteins, that lack about 40 amino acids, and analyzing poly(ADP-ribosyl)ated domain of p53.
We identified a GST-truncated p53 protein, in which about 40 amino acids are deleted in the DNA binding domain, that lacks the acceptor activity of poly(ADP-ribosyl)ation. To identify the poly(ADP-ribosyl)ated proteins in general, we made an affinity column that have the antibody against poly(ADP-ribose), 1OH. We are optimizing the condition to bind the proteins from centrosome fractions and from other cellular fractions. To prepare a large amount of proteins that are poly(ADP-ribosylated), we are collecting the proteins from Drosophila mutant that lacks poly(ADP-ribose) glycohydrolase.
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