PMID: 2505068Sep 1, 1989Paper

Cloning and expression of the Bacillus subtilis methyltransferase gene in Escherichia coli ada- cells

Mutation Research
K I KodamaM Sekiguchi

Abstract

DNA fragments of Bacillus subtilis were inserted into a plasmid vector that can multiply in Escherichia coli cells, and foreign genes were expressed under the control of the lac promoter. By selecting hybrid plasmids that confer an increased resistance to alkylating agents on E. coli ada- mutant cells, the B. subtilis gene dat, which encodes O6-methylguanine-DNA methyltransferase, was cloned. The Dat protein, with a molecular weight of about 20,000, could transfer the methyl group from methylated DNA to its own protein molecule. Based on the nucleotide sequence of the gene, it was deduced that the protein comprises 165 amino acids and that the molecular weight is 18,779. The presumptive amino acid sequence of Dat protein is homologous to the sequences of the E. coli Ogt protein and the C-terminal half of the Ada protein, both of which carry O6-methylguanine-DNA methyltransferase activity. The pentaamino acid sequence Pro-Cys-His-Arg-Val, the cysteine residue of which is the methyl acceptor site in Ada protein, was conserved in the 3 methyltransferase proteins. The structural similarity of these methyltransferases suggests possible evolution from a single ancestral gene.

References

Mar 27, 1979·Molecular & General Genetics : MGG·Y Yamamoto, M Sekiguchi
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Jan 1, 1988·Annual Review of Biochemistry·T LindahlY Nakabeppu
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Citations

Feb 1, 1986·Molecular & General Genetics : MGG·F Morohoshi, N Munakata
Jan 1, 1996·Journal of Cancer Research and Clinical Oncology·M SekiguchiT Tuzuki
Nov 1, 1990·Mutation Research·A E Pegg
Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J ThomaleM F Rajewsky
Sep 25, 1990·Nucleic Acids Research·F MorohoshiN Munakata
Jan 1, 1992·Japanese Journal of Cancer Research : Gann·M FukuharaM Sekiguchi
Apr 1, 1992·Molecular Microbiology·L Samson
Dec 1, 1996·Critical Reviews in Biochemistry and Molecular Biology·A Memisoglu, L Samson

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