Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1990 Sep 11;18(17):5051–5053. doi: 10.1093/nar/18.17.5051

The repair patch of E. coli (A)BC excinuclease.

Sibghat-Ullah 1, A Sancar 1, J E Hearst 1
PMCID: PMC332122  PMID: 2205836

Abstract

The size of repair patch made by E. coli DNA polymerase I (Poll) following the removal of a thymine-psoralen monoadduct by E. coli (A)BC excinuclease was determined by using an M13mp19 DNA with a single psoralen monoadduct at the polylinker region. Incubation of this substrate with (A)BC excinuclease, Poll and a combination of 3 dnTP plus 1 dNTP(alpha S) for each nucleotide, and DNA ligase resulted in a repair patch with phosphorothioate linkages. The preferential hydrolysis of phosphorothioate bonds by heating in iodoethanol revealed a patch size--with minimal nick translation--equal in length to the 12 nucleotide gap generated by this excision nuclease.

Full text

PDF
5051

Images in this article

5052Image
on p.5052

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ben-Ishai R., Sharon R. Patch size and base composition of ultraviolet light-induced repair synthesis in toluenized Escherichia coli. J Mol Biol. 1978 Apr 15;120(3):423–432. doi: 10.1016/0022-2836(78)90428-x. [DOI] [PubMed] [Google Scholar]
  2. Carlson K. M., Smith K. C. Effect of the uvrD3 mutation on ultraviolet radiation-induced DNA-repair replication in Escherichia coli K12. Mutat Res. 1981 Dec;84(2):257–262. doi: 10.1016/0027-5107(81)90195-0. [DOI] [PubMed] [Google Scholar]
  3. Caron P. R., Kushner S. R., Grossman L. Involvement of helicase II (uvrD gene product) and DNA polymerase I in excision mediated by the uvrABC protein complex. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4925–4929. doi: 10.1073/pnas.82.15.4925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cooper P. K., Hanawalt P. C. Heterogeneity of patch size in repair replicated DNA in Escherichia coli. J Mol Biol. 1972 Jun 14;67(1):1–10. doi: 10.1016/0022-2836(72)90381-6. [DOI] [PubMed] [Google Scholar]
  5. Gish G., Eckstein F. DNA and RNA sequence determination based on phosphorothioate chemistry. Science. 1988 Jun 10;240(4858):1520–1522. doi: 10.1126/science.2453926. [DOI] [PubMed] [Google Scholar]
  6. Kodadek T., Gamper H. Efficient synthesis of a supercoiled M13 DNA molecule containing a site specifically placed psoralen adduct and its use as a substrate for DNA replication. Biochemistry. 1988 May 3;27(9):3210–3215. doi: 10.1021/bi00409a013. [DOI] [PubMed] [Google Scholar]
  7. Ley R. D., Setlow R. B. Repair replication in Escherichia coli as measured by the photolysis of bromodeoxyuridine. Biophys J. 1972 Apr;12(4):420–431. doi: 10.1016/S0006-3495(72)86094-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nakamaye K. L., Gish G., Eckstein F., Vosberg H. P. Direct sequencing of polymerase chain reaction amplified DNA fragments through the incorporation of deoxynucleoside alpha-thiotriphosphates. Nucleic Acids Res. 1988 Nov 11;16(21):9947–9959. doi: 10.1093/nar/16.21.9947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rothman R. H. Dimer excision and repair replication patch size in recL152 mutant of Escherichia coli K-12. J Bacteriol. 1978 Oct;136(1):444–448. doi: 10.1128/jb.136.1.444-448.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rothman R. H. Dimer excision in Escherichia coli in the presence of caffeine. J Bacteriol. 1980 Jul;143(1):520–524. doi: 10.1128/jb.143.1.520-524.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sancar A., Rupp W. D. A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell. 1983 May;33(1):249–260. doi: 10.1016/0092-8674(83)90354-9. [DOI] [PubMed] [Google Scholar]
  12. Sancar A., Sancar G. B. DNA repair enzymes. Annu Rev Biochem. 1988;57:29–67. doi: 10.1146/annurev.bi.57.070188.000333. [DOI] [PubMed] [Google Scholar]
  13. Thomas D. C., Levy M., Sancar A. Amplification and purification of UvrA, UvrB, and UvrC proteins of Escherichia coli. J Biol Chem. 1985 Aug 15;260(17):9875–9883. [PubMed] [Google Scholar]
  14. Van Houten B., Gamper H., Hearst J. E., Sancar A. Analysis of sequential steps of nucleotide excision repair in Escherichia coli using synthetic substrates containing single psoralen adducts. J Biol Chem. 1988 Nov 15;263(32):16553–16560. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES