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. 1976 Nov;73(11):3979–3983. doi: 10.1073/pnas.73.11.3979

Identification and radiochemical purification of the recA protein of Escherichia coli K-12.

K McEntee, J E Hesse, W Epstein
PMCID: PMC431291  PMID: 792880

Abstract

The product of the recA gene of E. coli has been identified by labeling proteins synthesized in UV-treated cells after infection with specialized transducing phages carrying the recA gene. Following infection of UV-treated cells by lambda precA, which carries the recA+ gene, a major protein with a molecular weight of 43,000 is detected on polyacrylamide gels containing sodium dodecyl sulfate. This protein is also made after infection of suppressing hosts by lambda precA99, which carries an amber recA- mutation, but is not synthesized after infection of nonsuppressing hosts by this transducing phage. A spontaneous recatrevertant of lambda preca99 induces synthesis of this protein after infection of a nonsuppressing host. The product of the recA gene is a soluble protein found in a complex with a molecular weight of approximately 150,000 after mild detergent lysis of cells.

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Selected References

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

  1. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Brooks K., Clark A. J. Behavior of lambda bacteriophage in a recombination deficienct strain of Escherichia coli. J Virol. 1967 Apr;1(2):283–293. doi: 10.1128/jvi.1.2.283-293.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CLARK A. J., MARGULIES A. D. ISOLATION AND CHARACTERIZATION OF RECOMBINATION-DEFICIENT MUTANTS OF ESCHERICHIA COLI K12. Proc Natl Acad Sci U S A. 1965 Feb;53:451–459. doi: 10.1073/pnas.53.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark A. J., Chamberlin M., Boyce R. P., Howard-Flanders P. Abnormal metabolic response to ultraviolet light of a recombination deficient mutant of Escherichia coli K12. J Mol Biol. 1966 Aug;19(2):442–454. doi: 10.1016/s0022-2836(66)80015-3. [DOI] [PubMed] [Google Scholar]
  7. Clark A. J. Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet. 1973;7:67–86. doi: 10.1146/annurev.ge.07.120173.000435. [DOI] [PubMed] [Google Scholar]
  8. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Epstein W., Kim B. S. Potassium transport loci in Escherichia coli K-12. J Bacteriol. 1971 Nov;108(2):639–644. doi: 10.1128/jb.108.2.639-644.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldmark P. J., Linn S. An endonuclease activity from Escherichia coli absent from certain rec- strains. Proc Natl Acad Sci U S A. 1970 Sep;67(1):434–441. doi: 10.1073/pnas.67.1.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldmark P. J., Linn S. Purification and properties of the recBC DNase of Escherichia coli K-12. J Biol Chem. 1972 Mar 25;247(6):1849–1860. [PubMed] [Google Scholar]
  12. Gudas L. J., Pardee A. B. DNA synthesis inhibition and the induction of protein X in Escherichia coli. J Mol Biol. 1976 Mar 15;101(4):459–477. doi: 10.1016/0022-2836(76)90240-0. [DOI] [PubMed] [Google Scholar]
  13. Gudas L. J. The induction of protein X in DNA repair and cell division mutants of Escherichia coli. J Mol Biol. 1976 Jul 5;104(3):567–587. doi: 10.1016/0022-2836(76)90121-2. [DOI] [PubMed] [Google Scholar]
  14. Horii Z., Clark A. J. Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K12: isolation and characterization of mutants. J Mol Biol. 1973 Oct 25;80(2):327–344. doi: 10.1016/0022-2836(73)90176-9. [DOI] [PubMed] [Google Scholar]
  15. Howard-Flanders P., Theriot L. Mutants of Escherichia coli K-12 defective in DNA repair and in genetic recombination. Genetics. 1966 Jun;53(6):1137–1150. doi: 10.1093/genetics/53.6.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Inouye M., Pardee A. B. Changes of membrane proteins and their relation to deoxyribonucleic acid synthesis and cell division of Escherichia coli. J Biol Chem. 1970 Nov 10;245(21):5813–5819. [PubMed] [Google Scholar]
  17. Jaskunas S. R., Lindahl L., Nomura M. Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature. 1975 Oct 9;257(5526):458–462. doi: 10.1038/257458a0. [DOI] [PubMed] [Google Scholar]
  18. Jenkins S. T., Bennett P. M. Effect of mutations in deoxyribonucleic acid repair pathways on the sensitivity of Escherichia coli K-12 strains to nitrofurantoin. J Bacteriol. 1976 Mar;125(3):1214–1216. doi: 10.1128/jb.125.3.1214-1216.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kushner S. R., Nagaishi H., Clark A. J. Isolation of exonuclease VIII: the enzyme associated with sbcA indirect suppressor. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3593–3597. doi: 10.1073/pnas.71.9.3593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kushner S. R., Nagaishi H., Templin A., Clark A. J. Genetic recombination in Escherichia coli: the role of exonuclease I. Proc Natl Acad Sci U S A. 1971 Apr;68(4):824–827. doi: 10.1073/pnas.68.4.824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lloyd R. G., Low B., Godson G. N., Birge E. A. Isolation and characterization of an Escherichia coli K-12 mutant with a temperature-sensitive recA- phenotype. J Bacteriol. 1974 Oct;120(1):407–415. doi: 10.1128/jb.120.1.407-415.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Low R. L., Rashbaum S. A., Cozzarelli N. R. Purification and characterization of DNA polymerase III from Bacillus subtilis. J Biol Chem. 1976 Mar 10;251(5):1311–1325. [PubMed] [Google Scholar]
  23. McEntee K. Specialized transduction of recA by bacteriophage lambda. Virology. 1976 Mar;70(1):221–222. doi: 10.1016/0042-6822(76)90258-0. [DOI] [PubMed] [Google Scholar]
  24. Mount D. W. Isolation and genetic analysis of a strain of Escherichia coli K-12 with an amber recA mutation. J Bacteriol. 1971 Jul;107(1):388–389. doi: 10.1128/jb.107.1.388-389.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Neihardt F. C., Parker J., McKeever W. G. Function and regulation of aminoacyl-tRNA synthetases in prokaryotic and eukaryotic cells. Annu Rev Microbiol. 1975;29:215–250. doi: 10.1146/annurev.mi.29.100175.001243. [DOI] [PubMed] [Google Scholar]
  26. Ptashne M. ISOLATION OF THE lambda PHAGE REPRESSOR. Proc Natl Acad Sci U S A. 1967 Feb;57(2):306–313. doi: 10.1073/pnas.57.2.306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Witkin E. M. The mutability toward ultraviolet light of recombination-deficient strains of Escherichia coli. Mutat Res. 1969 Jul-Aug;8(1):9–14. doi: 10.1016/0027-5107(69)90135-3. [DOI] [PubMed] [Google Scholar]

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