1995 Fiscal Year Final Research Report Summary
Methylation and Demethylation of DNA in the Higher Animal
| Project/Area Number |
06680613
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional biochemistry
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| Research Institution | OSAKA UNIVERSITY |
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Principal Investigator |
TAJIMA Shoji Institute for Protein Research, associate professor, 蛋白質研究所, 助教授 (50132931)
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| Project Period (FY) |
1994 – 1995
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| Keywords | DNA methylation / DNA methyltransferase / DNA binding protein / Myogenesis / MyoD |
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| Research Abstract |
In the project, I aimed to shed light on the regulation of the DNA methylation of chromosomal DNA in vertebrate. For this purpose, I engaged in the following experiments and got the results as follows.
1. I have isolated DNA methyltransferase (MTase) cDNAs from chiken (avian) and Xenopus laevis (amphibian) and determined the nucleotide sequences. Both cDNAs were expressed in cultivated cells and their properties were analyzes. Overexpressed mouse MTase showed higher de novo methylation activity than chicken MTase.
2. I have identified a novel DNA binding protein, MMBP-3, that specifically recognizes the methylated form of c-Myc binding motif. The DNA binding activity of the MMBP-3 is high in the proliferating cells and decreases when the cell proliferation is arrested.
3. Overexpression of mouse MTase in myoblasts accelerates the myotube formation. This is caused by direct induction of MyoD gene transcription under proliferating conditions. The expression of MyoD gene positively correlates with the methylation of specific CpG sequences near transcriptional start site.
4. I have isolated a novel cDNA named AZ1, of which expression is induced with demethylating reagent. The AZ1 protein is localized to the pre-acrosome of spermatocytes.
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