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Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli.

Publication ,  Journal Article
Pukkila, PJ; Peterson, J; Herman, G; Modrich, P; Meselson, M
Published in: Genetics
August 1983

Two methods were used in an attempt to increase the efficiency and strand selectivity of methyl-directed mismatch repair of bacteriophage lambda heteroduplexes in E. coli. Previous studies of such repair used lambda DNA that was only partially methylated as the source of methylated chains. Also, transfection was carried out in methylating strains. Either of these factors might have been responsible for the incompleteness of the strand selectivity observed previously. In the first approach to increasing strand selectivity, heteroduplexes were transfected into a host deficient in methylation, but no changes in repair frequencies were observed. In the second approach, heteroduplexes were prepared using DNA that had been highly methylated in vitro with purified DNA adenine methylase as the source of methylated chains. In heteroduplexes having a repairable cI/+ mismatch, strand selectivity was indeed enhanced. In heteroduplexes with one chain highly methylated and the complementary chain unmethylated, the frequency of repair on the unmethylated chain increased to nearly 100%. Heteroduplexes with both chains highly methylated were not repaired at a detectable frequency. Thus, chains highly methylated by DNA adenine methylase were refractory to mismatch repair by this system, regardless of the methylation of the complementary chain. These results support the hypothesis that methyl-directed mismatch repair acts to correct errors of replication, thus lowering the mutation rate.

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Published In

Genetics

DOI

ISSN

0016-6731

Publication Date

August 1983

Volume

104

Issue

4

Start / End Page

571 / 582

Location

United States

Related Subject Headings

  • Methylation
  • Escherichia coli
  • Developmental Biology
  • DNA, Bacterial
  • DNA Repair
  • Bacteriophage lambda
  • Adenine
  • 3105 Genetics
  • 3101 Biochemistry and cell biology
  • 0604 Genetics
 

Citation

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Pukkila, P. J., Peterson, J., Herman, G., Modrich, P., & Meselson, M. (1983). Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli. Genetics, 104(4), 571–582. https://doi.org/10.1093/genetics/104.4.571
Pukkila, P. J., J. Peterson, G. Herman, P. Modrich, and M. Meselson. “Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli.Genetics 104, no. 4 (August 1983): 571–82. https://doi.org/10.1093/genetics/104.4.571.
Pukkila PJ, Peterson J, Herman G, Modrich P, Meselson M. Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli. Genetics. 1983 Aug;104(4):571–82.
Pukkila, P. J., et al. “Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli.Genetics, vol. 104, no. 4, Aug. 1983, pp. 571–82. Pubmed, doi:10.1093/genetics/104.4.571.
Pukkila PJ, Peterson J, Herman G, Modrich P, Meselson M. Effects of high levels of DNA adenine methylation on methyl-directed mismatch repair in Escherichia coli. Genetics. 1983 Aug;104(4):571–582.

Published In

Genetics

DOI

ISSN

0016-6731

Publication Date

August 1983

Volume

104

Issue

4

Start / End Page

571 / 582

Location

United States

Related Subject Headings

  • Methylation
  • Escherichia coli
  • Developmental Biology
  • DNA, Bacterial
  • DNA Repair
  • Bacteriophage lambda
  • Adenine
  • 3105 Genetics
  • 3101 Biochemistry and cell biology
  • 0604 Genetics