2005 Fiscal Year Final Research Report Summary
The mechanisms to eliminate oxidized nucleotides and maintenance of genetic information
| Project/Area Number |
16570149
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | Biomolecular Engineering Research Institute |
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Principal Investigator |
ISHIBASHI Toru Biomolecular Engineering Research Institute, Department of Biomolecular Engineering, Principal investigator, 機能制御研究部, 主席研究員 (90369041)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Riyoko Fukuoka Dental College, assistant professor, 歯学部, 助手 (10140865)
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| Project Period (FY) |
2004 – 2005
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| Keywords | oxidation / nucleotides / NUDT5 / MutT / 8-oxo-guanine |
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| Research Abstract |
To investigate the mechanism for cleaning up the nucleotide pools, we analyzed the enzymatic properties of MutT, NUDT5, and MTH1.
1. It was revealed that both MutT and MTH1 eliminate oxidized nucleoside diphosphates from the nucleotide pools. This indicates that the quality control of nucleoside diphosphates is important beyond the organisms. To understand the mechanisms further, we examined how the enzyme degrades the oxidized nucleoside triphosphates by using a-32P-labeled 8-oxo-GTP. Both MutT and MTH1 cleaved the phosphodiester bond between a and l phosphates, producing nucleoside monophosphates directly. Although 8-oxo-nuclsoside diphosphate can be used for DNA and RNA synthesis, 8-oxo-nucleoside monophosphaate never be phospholylated. These results are important to understand the cleaning up mechanisms of of nucleotide pools in organisms.
2. We prepared the polyclonal antibodies for NUDT5 and investigated the expression of NUDT5 in mouse organisms. NUDT5 was expressed in thymus, spleen, liver, heart and lung. And also, NUDT5 was predominantly expressed in cytoplasm. Using these antibodies, we isolated the protein complexes from HeLa cells and E.coli cells and performed Western blot analysis. These results indicated that both MutT and MUDT5 to form protein complex in a cell.
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