| Project/Area Number |
17370058
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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 |
Biophysics
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| Research Institution | Yokohama City University |
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Principal Investigator |
NISHIMURA Yoshifumi Yokohama City University, Division of Structural Biology, International Graduate School of Arts and Sciences, Professor (70107390)
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| Co-Investigator(Kenkyū-buntansha) |
AKASHI Satoko Yokohama City University, Division of Structural Biology, International Graduate School of Arts and Sciences, Associate Professor (10280728)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥14,050,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2007: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | Nucleic acid / Genome / Protein / Regulation of gene expression / Aging / NMR / General transcription factor / Structural biology / タンパク質 / 転写調節 |
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| Research Abstract |
We determined the structures of both free hTFIIEalpha C-tearminal acidic domain (AC-D) and its form bound to the N-terminal pleckstrin homology domain (PH-D) of the p62 subunit of hTFIIH. Intriguingly, hTFIIEalpha AC-D was found to share its binding surface on p62 PH-D with the acidic transactivation peptide fragment of the human tumor supressor protein p53. However, hTFIIEalpha AC-D employs an entirely novel binding mode, which differs from the amphipathic helix method used by many transcriptional activators. The binding surface between p62 PH-D and hTFIIEalpha AC-D is much broader than the specific binding surface between p62 PH-D and the p53 peptide fragment. Given the recent demonstration of TFIIE and TFIIH functions in post initiation events (i.e. promoter clearance) these data are of special relevance to the regulatory mechanisms involved.
Essential Sas-related acetyltransferase 1 (Esa1) contains a presumed chromodomain in addition to a histone acetyltransferase (HAT) domain. We initially determined the solution structure of the Esa1 presumed chromodomain, and showed it to consist of a well folded structure similar to the tudor domain. The domain showed no RNA/DNA binding ability. Since the N-terminus of the protein forms a helical turn, we prepared an N-terminally extended construct, which was surprisingly found to bind to RNA and to be critical for in vivo function. This extended protein contains an additional β-sheet which acts as a knot for the tudor domain. The knot does not cause a global change in the core structure but induces a well defined loop in the tudor domain itself, which is responsible for RNA binding. We made point mutants in an esa1 mutant gene in yeast and their functional abilities examined. The knotted tudor domain mutations which were lethal to the yeast lost RNA binding ability.
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