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. 1984 Feb;157(2):490–497. doi: 10.1128/jb.157.2.490-497.1984

Mutations in the DNA gyrB gene that are temperature sensitive for lambda site-specific recombination, Mu growth, and plasmid maintenance.

D I Friedman, L C Plantefaber, E J Olson, D Carver, M H O'Dea, M Gellert
PMCID: PMC215274  PMID: 6319362

Abstract

We report the isolation of two mutations in the gyrB gene of Escherichia coli K12 obtained from an initial selection for resistance to coumermycin A1 and a subsequent screening for bacteria that fail to support site-specific recombination of phage lambda, i.e., Him-. These two mutations have a temperature-sensitive Him- phenotype, supporting site-specific recombination efficiently at low temperature, but inefficiently at high temperatures. Like other Him mutants, the gyrB-him mutants fail to plate phage Mu; again this defect is observed only at high temperatures. Additional thermally sensitive characteristics have also been observed; growth of lambda as well as maintenance of the plasmids pBR322 and F' gal are reduced at high temperature. Restriction of foreign DNA imposed by a P1 prophage is also reduced in these mutants. The temperature-sensitive phenotypic characteristics imposed by both the gyrB-him-230(Ts) and gyrB-him-231(Ts) mutations correlate with in vitro studies that show decreased gyrase activity, especially at higher temperatures, and in vivo studies showing reduced supercoiling of lambda DNA in the mutants at high temperature.

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Selected References

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

  1. Abremski K., Gottesman S. The form of the DNA substrate required for excisive recombination of bacteriophage lambda. J Mol Biol. 1979 Jul 5;131(3):637–649. doi: 10.1016/0022-2836(79)90012-3. [DOI] [PubMed] [Google Scholar]
  2. Austin S., Ziese M., Sternberg N. A novel role for site-specific recombination in maintenance of bacterial replicons. Cell. 1981 Sep;25(3):729–736. doi: 10.1016/0092-8674(81)90180-x. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bolivar F., Rodriguez R. L., Betlach M. C., Boyer H. W. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene. 1977;2(2):75–93. doi: 10.1016/0378-1119(77)90074-9. [DOI] [PubMed] [Google Scholar]
  5. Botchan P. An electron microscopic comparison of transcription on linear and superhelical DNA. J Mol Biol. 1976 Jul 25;105(1):161–176. doi: 10.1016/0022-2836(76)90201-1. [DOI] [PubMed] [Google Scholar]
  6. Botchan P., Wang J. C., Echols H. Effect of circularity and superhelicity on transcription from bacteriophagelambda DNA. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3077–3081. doi: 10.1073/pnas.70.11.3077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cerdá-Olmedo E., Hanawalt P. C. The replication of the Escherichia coli chromosome studied by sequential nitrosoguanidine mutagenesis. Cold Spring Harb Symp Quant Biol. 1968;33:599–607. doi: 10.1101/sqb.1968.033.01.066. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Enquist L. W., Weisberg R. A. The red plaque test: a rapid method for identification of excision defective variants of bacteriophage lambda. Virology. 1976 Jul 1;72(1):147–153. doi: 10.1016/0042-6822(76)90319-6. [DOI] [PubMed] [Google Scholar]
  11. Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. doi: 10.1146/annurev.bi.50.070181.004311. [DOI] [PubMed] [Google Scholar]
  12. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gellert M., Mizuuchi K., O'Dea M. H., Nash H. A. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. doi: 10.1073/pnas.73.11.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gellert M., O'Dea M. H., Itoh T., Tomizawa J. Novobiocin and coumermycin inhibit DNA supercoiling catalyzed by DNA gyrase. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4474–4478. doi: 10.1073/pnas.73.12.4474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  16. Guarneros G., Echols H. New mutants of bacteriophage lambda with a specific defect in excision from the host chromosome. J Mol Biol. 1970 Feb 14;47(3):565–574. doi: 10.1016/0022-2836(70)90323-2. [DOI] [PubMed] [Google Scholar]
  17. Hayashi Y., Hayashi M. Template activities of the phi X-174 replicative allomorphic deoxyribonucleic acids. Biochemistry. 1971 Nov;10(23):4212–4218. doi: 10.1021/bi00799a009. [DOI] [PubMed] [Google Scholar]
  18. Hays J. B., Boehmer S. Antagonists of DNA gyrase inhibit repair and recombination of UV-irradiated phage lambda. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4125–4129. doi: 10.1073/pnas.75.9.4125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heilmann H., Burkardt H. J., Pühler A., Reeve J. N. Transposon mutagenesis of the gene encoding the bacteriophage P1 restriction endonuclease. Co-linearity of the gene and gene product. J Mol Biol. 1980 Dec 15;144(3):387–396. doi: 10.1016/0022-2836(80)90097-2. [DOI] [PubMed] [Google Scholar]
  20. Ikeda H., Tomizawa J. Prophage P1, and extrachromosomal replication unit. Cold Spring Harb Symp Quant Biol. 1968;33:791–798. doi: 10.1101/sqb.1968.033.01.091. [DOI] [PubMed] [Google Scholar]
  21. KAISER A. D., JACOB F. Recombination between related temperate bacteriophages and the genetic control of immunity and prophage localization. Virology. 1957 Dec;4(3):509–521. doi: 10.1016/0042-6822(57)90083-1. [DOI] [PubMed] [Google Scholar]
  22. Kikuchi Y., Nash H. Integrative recombination of bacteriophage lambda: requirement for supertwisted DNA in vivo and characterization of int. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1099–1109. doi: 10.1101/sqb.1979.043.01.122. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Kourilsky P., Perricaudet M., Gros D., Garapin A., Gottesman M., Fritsch A., Tiollais P. Description and properties of bacteriophage lambda vectors useful for the cloning of EcoRI DNA fragments. Biochimie. 1978;60(2):183–187. doi: 10.1016/s0300-9084(78)80752-4. [DOI] [PubMed] [Google Scholar]
  25. Kubo M., Kano Y., Nakamura H., Nagata A., Imamoto F. In vivo enhancement of general and specific transcription in Escherichia coli by DNA gyrase activity. Gene. 1979 Oct;7(2):153–171. doi: 10.1016/0378-1119(79)90030-1. [DOI] [PubMed] [Google Scholar]
  26. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  27. LIEB M. The establishment of lysogenicity in Escherichia coli. J Bacteriol. 1953 Jun;65(6):642–651. doi: 10.1128/jb.65.6.642-651.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Manly K. F., Signer E. R., Radding C. M. Nonessential functions of bacteriophage lambda. Virology. 1969 Feb;37(2):177–188. doi: 10.1016/0042-6822(69)90197-4. [DOI] [PubMed] [Google Scholar]
  29. McHugh G. L., Swartz M. N. Elimination of plasmids from several bacterial species by novobiocin. Antimicrob Agents Chemother. 1977 Sep;12(3):423–426. doi: 10.1128/aac.12.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Menzel R., Gellert M. Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Cell. 1983 Aug;34(1):105–113. doi: 10.1016/0092-8674(83)90140-x. [DOI] [PubMed] [Google Scholar]
  31. Miller H. I., Friedman D. I. An E. coli gene product required for lambda site-specific recombination. Cell. 1980 Jul;20(3):711–719. doi: 10.1016/0092-8674(80)90317-7. [DOI] [PubMed] [Google Scholar]
  32. Miller H. I., Kikuchi A., Nash H. A., Weisberg R. A., Friedman D. I. Site-specific recombination of bacteriophage lambda: the role of host gene products. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1121–1126. doi: 10.1101/sqb.1979.043.01.125. [DOI] [PubMed] [Google Scholar]
  33. Mizuuchi K., Gellert M., Nash H. A. Involement of supertwisted DNA in integrative recombination of bacteriophage lambda. J Mol Biol. 1978 May 25;121(3):375–392. doi: 10.1016/0022-2836(78)90370-4. [DOI] [PubMed] [Google Scholar]
  34. Mizuuchi K., Mizuuchi M. Integrative recombination of bacteriophage lambda: in vitro study of the intermolecular reaction. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1111–1114. doi: 10.1101/sqb.1979.043.01.123. [DOI] [PubMed] [Google Scholar]
  35. Modrich P., Zabel D. EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme. J Biol Chem. 1976 Oct 10;251(19):5866–5874. [PubMed] [Google Scholar]
  36. Nash H. A. Integration and excision of bacteriophage lambda: the mechanism of conservation site specific recombination. Annu Rev Genet. 1981;15:143–167. doi: 10.1146/annurev.ge.15.120181.001043. [DOI] [PubMed] [Google Scholar]
  37. Nash H. A. Integrative recombination in bacteriophage lambda: analysis of recombinant DNA. J Mol Biol. 1975 Feb 5;91(4):501–514. doi: 10.1016/0022-2836(75)90276-4. [DOI] [PubMed] [Google Scholar]
  38. Nash H. A. LambdaattB-attP, a lambda derivative containing both sites involved in integrative recombination. Virology. 1974 Jan;57(1):207–216. doi: 10.1016/0042-6822(74)90121-4. [DOI] [PubMed] [Google Scholar]
  39. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  40. Ryan M. J. Coumermycin A1: A preferential inhibitor of replicative DNA synthesis in Escherichia coli. I. In vivo characterization. Biochemistry. 1976 Aug 24;15(17):3769–3777. doi: 10.1021/bi00662a020. [DOI] [PubMed] [Google Scholar]
  41. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol. 1982 Dec 25;162(4):729–773. doi: 10.1016/0022-2836(82)90546-0. [DOI] [PubMed] [Google Scholar]
  42. Sanzey B. Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase. J Bacteriol. 1979 Apr;138(1):40–47. doi: 10.1128/jb.138.1.40-47.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Shuman H., Schwartz M. The effect of nalidixic acid on the expression of some genes in Escherichia coli K-12. Biochem Biophys Res Commun. 1975 May 5;64(1):204–209. doi: 10.1016/0006-291x(75)90239-9. [DOI] [PubMed] [Google Scholar]
  44. Smith C. L., Kubo M., Imamoto F. Promoter-specific inhibition of transcription by antibiotics which act on DNA gyrase. Nature. 1978 Oct 5;275(5679):420–423. doi: 10.1038/275420a0. [DOI] [PubMed] [Google Scholar]
  45. Sternberg N. A characterization of bacteriophage P1 DNA fragments cloned in a lambda vector. Virology. 1979 Jul 15;96(1):129–142. doi: 10.1016/0042-6822(79)90179-x. [DOI] [PubMed] [Google Scholar]
  46. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Taylor D. E., Levine J. G. Characterization of a plasmid mutation affecting maintenance, transfer and elimination by novobiocin. Mol Gen Genet. 1979 Jul 13;174(2):127–133. doi: 10.1007/BF00268350. [DOI] [PubMed] [Google Scholar]
  48. Wolfson J. S., Hooper D. C., Swartz M. N., McHugh G. L. Antagonism of the B subunit of DNA gyrase eliminates plasmids pBR322 and pMG110 from Escherichia coli. J Bacteriol. 1982 Oct;152(1):338–344. doi: 10.1128/jb.152.1.338-344.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yoshida R. K., Miller J. L., Miller H. I., Friedman D. I., Howe M. M. Isolation and mapping of Mu nu mutants which grow in him mutants of E. coli. Virology. 1982 Jul 15;120(1):269–272. doi: 10.1016/0042-6822(82)90027-7. [DOI] [PubMed] [Google Scholar]
  50. ZICHICHI M. L., KELLENBERGER G. Two distinct functions in the lysogenization process: the repression of phage multiplication and incorporation of the prophage in the bacterial genome. Virology. 1963 Apr;19:450–460. doi: 10.1016/0042-6822(63)90038-2. [DOI] [PubMed] [Google Scholar]

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