1989 Fiscal Year Final Research Report Summary
Regulation of Gene Expression by (ADP-ribosyl)ation Reaction
| Project/Area Number |
63480136
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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 |
Pathological medical chemistry
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| Research Institution | Kochi Medical School |
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Principal Investigator |
SHIZUTA Yutaka Kochi Medical School, Professor., 医学部, 教授 (50025631)
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| Co-Investigator(Kenkyū-buntansha) |
TODA Katsumi Kochi Medical School, Associate Professor., 医学部, 助教授 (40197893)
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| Project Period (FY) |
1988 – 1989
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| Keywords | ADP-ribosylation / cDNA cloning / Genomic DNA cloning / Monoclonal Antibody / Microinjection |
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| Research Abstract |
ADP-ribose transferase plays an important role in gene expression of eukaryotic cells. In the present study, we have isolated CDNA as well as genomic DNA for the enzyme and elucidated the characteristics of the gene as follows.
1) The open reading frame of the cDNA encodes a protein of 1,0141 amino acid residues with a molecular weight of 113,153. The DNA binding domain involves a homologus repeat in the sequence and exhibits a sequence homology with localized region of transforming proteins such as c-fos and v-fos.
2) The promoter region of the transferase gene lacks TATA box and CAT box, but it contains two potent Sp1 binding sites and three putative AP-2 binding elements. Northern blot analysis has revealed that transcription of the transferase gene is markedly stimulated both by cyclic AMP and by phorbol ester.
3) Using domain-specific nionoclonal antibodies, the half life of the transferase in the living cells was determiner to be approximately 15 hours. The antibodies appear to inhibit thymidine uncorporation into living cells, suggesting that the enzyme also plays an important role in DNA replication.
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