Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jan;173(1):150–155. doi: 10.1128/jb.173.1.150-155.1991

Isolation of temperature-sensitive McrA and McrB mutations and complementation analysis of the McrBC region of Escherichia coli K-12.

A Piekarowicz 1, R Yuan 1, D C Stein 1
PMCID: PMC207168  PMID: 1987114

Abstract

We isolated temperature-sensitive mcrA and mcrBC mutants of Escherichia coli. At 42 degrees C, they were unable to restrict the T-even bacteriophages T6gt and T4gt or plasmids encoding cloned DNA methylase genes whose specificities confer sensitivity to the McrA and McrBC nucleases. Complementation analysis of the McrBC region (mcrB251) with the complete cloned McrBC system or a derivative with mcrB alone indicated that the mutation shows an absolute defect for the restriction of DNA containing hydroxymethylcytosine and a thermosensitive defect for the restriction of DNA containing methylcytosine. The properties of the McrA temperature-sensitive mutants suggest that some of these mutations can also influence the restriction of DNA containing hydroxymethylcytosine or methylcytosine residues.

Full text

PDF
153

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blumenthal R. M., Gregory S. A., Cooperider J. S. Cloning of a restriction-modification system from Proteus vulgaris and its use in analyzing a methylase-sensitive phenotype in Escherichia coli. J Bacteriol. 1985 Nov;164(2):501–509. doi: 10.1128/jb.164.2.501-509.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dila D., Sutherland E., Moran L., Slatko B., Raleigh E. A. Genetic and sequence organization of the mcrBC locus of Escherichia coli K-12. J Bacteriol. 1990 Sep;172(9):4888–4900. doi: 10.1128/jb.172.9.4888-4900.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Georgopoulos C. P. Isolation and preliminary characterization of T4 mutants with nonglucosylated DNA. Biochem Biophys Res Commun. 1967 Jul 21;28(2):179–184. doi: 10.1016/0006-291x(67)90426-3. [DOI] [PubMed] [Google Scholar]
  5. Greener A., Hill C. W. Identification of a novel genetic element in Escherichia coli K-12. J Bacteriol. 1980 Oct;144(1):312–321. doi: 10.1128/jb.144.1.312-321.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heitman J., Model P. Site-specific methylases induce the SOS DNA repair response in Escherichia coli. J Bacteriol. 1987 Jul;169(7):3243–3250. doi: 10.1128/jb.169.7.3243-3250.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kleckner N., Barker D. F., Ross D. G., Botstein D. Properties of the translocatable tetracycline-resistance element Tn10 in Escherichia coli and bacteriophage lambda. Genetics. 1978 Nov;90(3):427–461. doi: 10.1093/genetics/90.3.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LURIA S. E., HUMAN M. L. A nonhereditary, host-induced variation of bacterial viruses. J Bacteriol. 1952 Oct;64(4):557–569. doi: 10.1128/jb.64.4.557-569.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Noyer-Weidner M., Diaz R., Reiners L. Cytosine-specific DNA modification interferes with plasmid establishment in Escherichia coli K12: involvement of rglB. Mol Gen Genet. 1986 Dec;205(3):469–475. doi: 10.1007/BF00338084. [DOI] [PubMed] [Google Scholar]
  10. Piekarowicz A., Yuan R., Stein D. C. Construction of a temperature-sensitive mutation for the direct identification of plasmids encoding DNA methyltransferases. Gene. 1988 Dec 25;74(1):233–235. doi: 10.1016/0378-1119(88)90294-6. [DOI] [PubMed] [Google Scholar]
  11. REVEL H. R., HATTMAN S., LURIA S. E. MUTANTS OF BACTERIOPHAGES T2 AND T6 DEFECTIVE IN ALPHA-GLUCOSYL TRANSFERASE. Biochem Biophys Res Commun. 1965 Feb 17;18:545–550. doi: 10.1016/0006-291x(65)90788-6. [DOI] [PubMed] [Google Scholar]
  12. Raleigh E. A., Murray N. E., Revel H., Blumenthal R. M., Westaway D., Reith A. D., Rigby P. W., Elhai J., Hanahan D. McrA and McrB restriction phenotypes of some E. coli strains and implications for gene cloning. Nucleic Acids Res. 1988 Feb 25;16(4):1563–1575. doi: 10.1093/nar/16.4.1563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Raleigh E. A., Trimarchi R., Revel H. Genetic and physical mapping of the mcrA (rglA) and mcrB (rglB) loci of Escherichia coli K-12. Genetics. 1989 Jun;122(2):279–296. doi: 10.1093/genetics/122.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Raleigh E. A., Wilson G. Escherichia coli K-12 restricts DNA containing 5-methylcytosine. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9070–9074. doi: 10.1073/pnas.83.23.9070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Revel H. R. Restriction of nonglucosylated T-even bacteriophage: properties of permissive mutants of Escherichia coli B and K12. Virology. 1967 Apr;31(4):688–701. doi: 10.1016/0042-6822(67)90197-3. [DOI] [PubMed] [Google Scholar]
  16. Ross T. K., Achberger E. C., Braymer H. D. Characterization of the Escherichia coli modified cytosine restriction (mcrB) gene. Gene. 1987;61(3):277–289. doi: 10.1016/0378-1119(87)90191-0. [DOI] [PubMed] [Google Scholar]
  17. Ross T. K., Achberger E. C., Braymer H. D. Identification of a second polypeptide required for McrB restriction of 5-methylcytosine-containing DNA in Escherichia coli K12. Mol Gen Genet. 1989 Apr;216(2-3):402–407. doi: 10.1007/BF00334382. [DOI] [PubMed] [Google Scholar]
  18. Ross T. K., Achberger E. C., Braymer H. D. Nucleotide sequence of the McrB region of Escherichia coli K-12 and evidence for two independent translational initiation sites at the mcrB locus. J Bacteriol. 1989 Apr;171(4):1974–1981. doi: 10.1128/jb.171.4.1974-1981.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ross T. K., Braymer H. D. Localization of a genetic region involved in McrB restriction by Escherichia coli K-12. J Bacteriol. 1987 Apr;169(4):1757–1759. doi: 10.1128/jb.169.4.1757-1759.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sozhamannan S., Dharmalingam K. Molecular cloning and identification of protein products of rglB locus of Escherichia coli. Gene. 1988 Dec 25;74(1):51–52. doi: 10.1016/0378-1119(88)90249-1. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES