A gene required for very short patch repair in Escherichia coli is adjacent to the DNA cytosine methylase gene.
Deamination of 5-methylcytosine in DNA results in T/G mismatches. If unrepaired, these mismatches can lead to C-to-T transition mutations. The very short patch (VSP) repair process in Escherichia coli counteracts the mutagenic process by repairing the mismatches in favor of the G-containing strand. Previously we have shown that a plasmid containing an 11-kilobase fragment from the E. coli chromosome can complement a chromosomal mutation defective in both cytosine methylation and VSP repair. We have now mapped the regions essential for the two phenotypes. In the process, we have constructed plasmids that complement the chromosomal mutation for methylation, but not for repair, and vice versa. The genes responsible for these phenotypes have been identified by DNA sequence analysis. The gene essential for cytosine methylation, dcm, is predicted to code for a 473-amino-acid protein and is not required for VSP repair. It is similar to other DNA cytosine methylases and shares extensive sequence similarity with its isoschizomer, EcoRII methylase. The segment of DNA essential for VSP repair contains a gene that should code for a 156-amino-acid protein. This gene, named vsr, is not essential for DNA methylation. Remarkably, the 5' end of this gene appears to overlap the 3' end of dcm. The two genes appear to be transcribed from a common promoter but are in different translational registers. This gene arrangement may assure that Vsr is produced along with Dcm and may minimize the mutagenic effects of cytosine methylation.
Sohail
A
A
Cold Spring Harbor Laboratory, New York 11724.
Lieb
M
M
Dar
M
M
Bhagwat
A S
AS
D000595:Amino Acid Sequence; D001483:Base Sequence; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics*; D004260:DNA Repair; D015257:DNA-Cytosine Methylases / Q000235:genetics; D004706:Endodeoxyribonucleases / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D005798:Genes, Bacterial; D008040:Genetic Linkage; D008969:Molecular Sequence Data; D010641:Phenotype; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D029968:Escherichia coli Proteins; C029528:DNA modification methylase EcoRII; D015257:DNA-Cytosine Methylases; D004248:DNA (Cytosine-5-)-Methyltransferases; D004706:Endodeoxyribonucleases; C000596250:Vsr protein, E coli