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Methyl-directed repair of DNA base-pair mismatches in vitro.

Publication ,  Journal Article
Lu, AL; Clark, S; Modrich, P
Published in: Proc Natl Acad Sci U S A
August 1983

An assay has been developed that permits analysis of DNA mismatch repair in cell-free extracts of Escherichia coli. The method relies on repair of heteroduplex molecules of f1 R229 DNA, which contain a base-pair mismatch within the single EcoRI site of the molecule. As observed with mismatch heteroduplexes of lambda DNA [Pukkila, P. J., Peterson, J., Herman, G., Modrich, P. & Meselson, M. (1983) Genetics, in press], in vivo mismatch correction of f1 heteroduplexes is directed by the state of dam methylation of d(G-A-T-C) sequences within the DNA duplex. Thus, the heteroduplex (formula: see book) is repaired in vivo to an EcoRI-sensitive form if the strand bearing the wild-type EcoRI sequence carries the dam modification and the other does not. Such molecules are also subject to mismatch repair by E. coli extracts. The in vitro activity is also dependent on ATP, the state of dam methylation of mismatch heteroduplexes, and products of mutH, mutL, mutS, and uvrE loci. However, crude fractions deficient in these gene products do complement in the cell-free system, thus providing assays for their isolation. The in vitro reaction is accompanied by repair synthesis on the unmethylated DNA strand.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

August 1983

Volume

80

Issue

15

Start / End Page

4639 / 4643

Location

United States

Related Subject Headings

  • Species Specificity
  • Mutation
  • Genetic Complementation Test
  • Escherichia coli
  • Deoxyribonuclease EcoRI
  • DNA Restriction Enzymes
  • DNA Repair
  • Coliphages
  • Base Sequence
  • Base Composition
 

Citation

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Lu, A. L., Clark, S., & Modrich, P. (1983). Methyl-directed repair of DNA base-pair mismatches in vitro. Proc Natl Acad Sci U S A, 80(15), 4639–4643. https://doi.org/10.1073/pnas.80.15.4639
Lu, A. L., S. Clark, and P. Modrich. “Methyl-directed repair of DNA base-pair mismatches in vitro.Proc Natl Acad Sci U S A 80, no. 15 (August 1983): 4639–43. https://doi.org/10.1073/pnas.80.15.4639.
Lu AL, Clark S, Modrich P. Methyl-directed repair of DNA base-pair mismatches in vitro. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4639–43.
Lu, A. L., et al. “Methyl-directed repair of DNA base-pair mismatches in vitro.Proc Natl Acad Sci U S A, vol. 80, no. 15, Aug. 1983, pp. 4639–43. Pubmed, doi:10.1073/pnas.80.15.4639.
Lu AL, Clark S, Modrich P. Methyl-directed repair of DNA base-pair mismatches in vitro. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4639–4643.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

August 1983

Volume

80

Issue

15

Start / End Page

4639 / 4643

Location

United States

Related Subject Headings

  • Species Specificity
  • Mutation
  • Genetic Complementation Test
  • Escherichia coli
  • Deoxyribonuclease EcoRI
  • DNA Restriction Enzymes
  • DNA Repair
  • Coliphages
  • Base Sequence
  • Base Composition