Isolation of a Mutant of Escherichia coli Defective in Cytosine-Specific Deoxyribonucleic Acid Methylase Activity and in Partial Protection of Bacteriophage λ Against Restriction by Cells Containing the N-3 Drug-Resistance Factor
- 1 January 1973
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 115 (3) , 1103-1107
- https://doi.org/10.1128/jb.115.3.1103-1107.1973
Abstract
A mutant (designated mec−) of Escherichia coli F+ 100 endo I−su+ rK−mK+ has been isolated which is defective in cytosine-specific deoxyribonucleic acid (DNA) methylase activity. The DNA of this mutant, as well as the DNA of phages λ and fd propagated in it, is virtually devoid of 5-methyl-cytosine (MeC); in contrast, the mutation has no significant effect on the level of N6-methyladenine in DNA. Phage λ grown on the mec− mutant is more strongly restricted by N-3-containing cells than is λ grown on the mec+ parent. These results suggest that methylation of certain cytosine residues by the E. coli K-12 enzyme partially protects λ DNA from either the N-3 restriction nuclease or against secondary degradation subsequent to N-3-specific degradation. Analysis of the MeC level in viral and cellular DNA obtained from mec+, mec+ (mN3+), and mec− (mN3+) strains has led to the conclusion that the R-factor controlled DNA-cytosine methylase may be capable of methylating a sequence(s) which is a substrate for the K-12 enzyme.Keywords
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