Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Sep;78(9):5450–5454. doi: 10.1073/pnas.78.9.5450

Identification of the uvrC gene product.

A Sancar, B M Kacinski, D L Mott, W D Rupp
PMCID: PMC348763  PMID: 7029536

Abstract

We have constructed a multicopy plasmid that carries the uvrC gene of Escherichia coli. By inserting the transposon Tn1000 (previously designated gamma delta) into this plasmid, we obtained many derivatives that fail to complement uvrC34. The proteins synthesized by the original plasmid and the uvrC::Tn1000 derivatives were labeled in maxicells and analyzed on gels, demonstrating that a protein of Mr 70,000 encoded by the original uvrC+ plasmid was absent from the mutated noncomplementing derivatives; this protein is presumed to be the uvrC gene product. We found that this protein of Mr 70,000 binds to DNA and have partially purified the uvrC protein by DNA-cellulose chromatography. Because some of the uvrC::Tn1000 derivatives produce truncated polypeptides, the orientation of expression and the location of the promoter were determined by correlating the sizes of the truncated polypeptides with the sites of insertion of Tn1000.

Full text

PDF
5450

Images in this article

5451Image
on p.5451
5452Image
on p.5452
5453Image
on p.5453

Selected References

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

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  3. Gudas L. J., Pardee A. B. Model for regulation of Escherichia coli DNA repair functions. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2330–2334. doi: 10.1073/pnas.72.6.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  5. Sancar A., Clarke N. D., Griswold J., Kennedy W. J., Rupp W. D. Identification of the uvrB gene product. J Mol Biol. 1981 May 5;148(1):63–76. doi: 10.1016/0022-2836(81)90235-7. [DOI] [PubMed] [Google Scholar]
  6. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Sancar A., Rupp W. D. Cloning of uvrA, lexC and ssb genes of Escherichia coli. Biochem Biophys Res Commun. 1979 Sep 12;90(1):123–129. doi: 10.1016/0006-291x(79)91598-5. [DOI] [PubMed] [Google Scholar]
  8. Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
  9. Seeberg E., Nissen-Meyer J., Strike P. Incision of ultraviolet-irradiated DNA by extracts of E. coli requires three different gene products. Nature. 1976 Oct 7;263(5577):524–526. doi: 10.1038/263524a0. [DOI] [PubMed] [Google Scholar]
  10. Seeberg E. Reconstitution of an Escherichia coli repair endonuclease activity from the separated uvrA+ and uvrB+/uvrC+ gene products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2569–2573. doi: 10.1073/pnas.75.6.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Seeberg E., Rupp W. D., Strike P. Impaired incision of ultraviolet-irradiated deoxyribonucleic acid in uvrC mutants of Escherichia coli. J Bacteriol. 1980 Oct;144(1):97–104. doi: 10.1128/jb.144.1.97-104.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sharma S., Moses R. E. uvrC gene function in excision repair in toluene-treated Escherichia coli. J Bacteriol. 1979 Jan;137(1):397–408. doi: 10.1128/jb.137.1.397-408.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Silverman M., Matsumura P., Hilmen M., Simon M. Characterization of lambda Escherichia coli hybrids carrying chemotaxis genes. J Bacteriol. 1977 May;130(2):877–887. doi: 10.1128/jb.130.2.877-887.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Yoakum G. H., Grossman L. Identification of E. coli uvrC protein. Nature. 1981 Jul 9;292(5819):171–173. doi: 10.1038/292171a0. [DOI] [PubMed] [Google Scholar]
  15. Yoakum G. H., Kushner S. R., Grossman L. Isolation of plasmids carrying either the uvrC or uvrC uvrA and ssb genes of Escherichia coli K-12. Gene. 1980 Dec;12(3-4):243–248. doi: 10.1016/0378-1119(80)90106-7. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES