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KMID : 1161519980020020287
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Animal Cells and Systems
1998 Volume.2 No. 2 p.287 ~ p.295
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Analysis for regulatory elements in yeast MGMT gene transcription
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Joo Jae-Hoon
Kim Woo-Jae
Rho Jae-Kyun
Kim Jae-Hyun
Choe Soo-Young
Park Sang-Dai
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Abstract
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The Saccharomyces cerevisiae MGMT gene encodes a O6?methylguanine DNA methyltransferase that protects cells from mutation or death by DNA alkylating agents. Using an in vitro transcription system, we analyzed its promoter region to find regulatory elements for transcription initiation. DNase I foot print ing and a transcription assay showed that a functional TATA box, 5'?TGATATAGCA?3’, is located in the region spanning from ?25 to ?34. We also found one upstream repressing sequence (URS), ?333 to ?213, by promoter deletion and competition analysis. Gel mobility shift assays and Southwestern blot analysis using URS region indicate specific complex formations. These results indicate that several c/s?acting and frans?acting elements might be involved in the transcriptional regulation of the S. cerevisiae MGMT gene
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KEYWORD
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Saccharomyces cerevisiae, O6-methylguanine-DNA methyltransferase (MGMT), In vitro transcription, TATA box, Upstream repressing sequence (URS)
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