1992 Fiscal Year Final Research Report Summary
Regulation of DNA replication and transcription by poly (ADP-ribose)
| Project/Area Number |
03454548
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
代謝生物化学
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| Research Institution | Kochi Medical School |
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Principal Investigator |
SHIZUTA Yutaka Kochi Medical School Dept. Med. Chem. Professor, 医学部, 教授 (50025631)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAMOTO Takeshi Kochi Medical School Dept. Med. Chem. Research associate, 医学部, 助手 (50224861)
MIYAHARA Kaoru Kochi Medical School Dept. Med. Chem. Research associate, 医学部, 助手 (30229877)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Poly (ADP-ribose) / DNA polymerase / DNA replication / DNA transcription / Steroid hormones / cDNA |
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| Research Abstract |
Poly (ADP-ribose) polymerase, an enzyme localized in the nucleic of enkaryotic cells, catalyzes the polymerization of the ADP ribose moisty of NAD to form poly (ADP-ribose) which is cavalently bound to various nuclear proteins. To elucidate the physiological function of the enzyme, we attempted to elucidate the characteristics of the promoter region of the gene for this enzyme and to characterized the role of this protein in DNA replication in vitro.
1) The promoter region contains two Sp 1 binding sites and three putative AP 2 binding elements. The promoter activity is enhanced both by cAMP and phorbol ester as judged by CAT assay and by Northern blot analysis.
2) Poly (ADP-ribose) polymerase stimulates DNA polymerase alpha by about 6 to 60 fold in a dose-depentent manner. It has no effect on the activities of DNA polymerase beta,gamma,and primase. Poly (ADP-ribose) polymerase is coimmunoprecipitaed with DNA polymerase alpha using anti-DNA polymerase antibody, indicating that poly (ADP-ribose) polymerase is physically associated with DNA polymerase alpha.
3) Steroid hormones affect level of poly (ADP-ribose) in various tissues. In relation to the above findings, several cDNAs and genomic DNAs for steroid hormone synthetase such as cortisol synthetase (P-45011beta), aldosterone synthase (P-450C18), and estrogen synthase (P-450arom) have been cloned and sequenced. Using CAT assay, regulation of their genes has been investigated in detail.
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