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. 1993 Jun;175(12):3798–3811. doi: 10.1128/jb.175.12.3798-3811.1993

Involvement of Escherichia coli FIS protein in maintenance of bacteriophage mu lysogeny by the repressor: control of early transcription and inhibition of transposition.

M Bétermier 1, I Poquet 1, R Alazard 1, M Chandler 1
PMCID: PMC204797  PMID: 8389742

Abstract

The Escherichia coli FIS (factor for inversion stimulation) protein has been implicated in assisting bacteriophage Mu repressor, c, in maintaining the lysogenic state under certain conditions. In a fis strain, a temperature-inducible Mucts62 prophage is induced at lower temperatures than in a wild-type host (M. Bétermier, V. Lefrère, C. Koch, R. Alazard, and M. Chandler, Mol. Microbiol. 3:459-468, 1989). Increasing the prophage copy number rendered Mucts62 less sensitive to this effect of the fis mutation, which thus seems to depend critically on the level of repressor activity. The present study also provides evidence that FIS affects the control of Mu gene expression and transposition. As judged by the use of lac transcriptional fusions, repression of early transcription was reduced three- to fourfold in a fis background, and this could be compensated by an increase in cts62 gene copy number. c was also shown to inhibit Mu transposition two- to fourfold less strongly in a fis host. These modulatory effects, however, could not be correlated to sequence-specific binding of FIS to the Mu genome, in particular to the strong site previously identified on the left end. We therefore speculate that a more general function of FIS is responsible for the observed modulation of Mu lysogeny.

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Selected References

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

  1. Alazard R., Bétermier M., Chandler M. Escherichia coli integration host factor stabilizes bacteriophage Mu repressor interactions with operator DNA in vitro. Mol Microbiol. 1992 Jun;6(12):1707–1714. doi: 10.1111/j.1365-2958.1992.tb00895.x. [DOI] [PubMed] [Google Scholar]
  2. Ball C. A., Johnson R. C. Multiple effects of Fis on integration and the control of lysogeny in phage lambda. J Bacteriol. 1991 Jul;173(13):4032–4038. doi: 10.1128/jb.173.13.4032-4038.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Bétermier M., Lefrère V., Koch C., Alazard R., Chandler M. The Escherichia coli protein, Fis: specific binding to the ends of phage Mu DNA and modulation of phage growth. Mol Microbiol. 1989 Apr;3(4):459–468. doi: 10.1111/j.1365-2958.1989.tb00192.x. [DOI] [PubMed] [Google Scholar]
  5. Cam K., Béjar S., Gil D., Bouché J. P. Identification and sequence of gene dicB: translation of the division inhibitor from an in-phase internal start. Nucleic Acids Res. 1988 Jul 25;16(14A):6327–6338. doi: 10.1093/nar/16.14.6327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chandler M., Galas D. J. Cointegrate formation mediated by Tn9. II. Activity of IS1 is modulated by external DNA sequences. J Mol Biol. 1983 Oct 15;170(1):61–91. doi: 10.1016/s0022-2836(83)80227-7. [DOI] [PubMed] [Google Scholar]
  7. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Colonna B., Hofnung M. rho Mutations restore lamB expression in E. coli K12 strains with an inactive malB region. Mol Gen Genet. 1981;184(3):479–483. doi: 10.1007/BF00352526. [DOI] [PubMed] [Google Scholar]
  9. Coulondre C., Miller J. H. Genetic studies of the lac repressor. III. Additional correlation of mutational sites with specific amino acid residues. J Mol Biol. 1977 Dec 15;117(3):525–567. doi: 10.1016/0022-2836(77)90056-0. [DOI] [PubMed] [Google Scholar]
  10. Craigie R., Arndt-Jovin D. J., Mizuuchi K. A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: protein and DNA substrate requirements. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7570–7574. doi: 10.1073/pnas.82.22.7570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Craigie R., Mizuuchi M., Mizuuchi K. Site-specific recognition of the bacteriophage Mu ends by the Mu A protein. Cell. 1984 Dec;39(2 Pt 1):387–394. doi: 10.1016/0092-8674(84)90017-5. [DOI] [PubMed] [Google Scholar]
  12. Drlica K., Rouviere-Yaniv J. Histonelike proteins of bacteria. Microbiol Rev. 1987 Sep;51(3):301–319. doi: 10.1128/mr.51.3.301-319.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Faelen M., Toussaint A., De Lafonteyne J. Model for the enchancement of lambde-gal integration into partially induced Mu-1 lysogens. J Bacteriol. 1975 Mar;121(3):873–882. doi: 10.1128/jb.121.3.873-882.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Falconi M., McGovern V., Gualerzi C., Hillyard D., Higgins N. P. Mutations altering chromosomal protein H-NS induce mini-Mu transposition. New Biol. 1991 Jun;3(6):615–625. [PubMed] [Google Scholar]
  15. Fayet O., Louarn J. M., Georgopoulos C. Suppression of the Escherichia coli dnaA46 mutation by amplification of the groES and groEL genes. Mol Gen Genet. 1986 Mar;202(3):435–445. doi: 10.1007/BF00333274. [DOI] [PubMed] [Google Scholar]
  16. Fellay R., Frey J., Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. doi: 10.1016/0378-1119(87)90041-2. [DOI] [PubMed] [Google Scholar]
  17. Filutowicz M., Ross W., Wild J., Gourse R. L. Involvement of Fis protein in replication of the Escherichia coli chromosome. J Bacteriol. 1992 Jan;174(2):398–407. doi: 10.1128/jb.174.2.398-407.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Friedman D. I. Integration host factor: a protein for all reasons. Cell. 1988 Nov 18;55(4):545–554. doi: 10.1016/0092-8674(88)90213-9. [DOI] [PubMed] [Google Scholar]
  19. Gama M. J., Toussaint A., Higgins N. P. Stabilization of bacteriophage Mu repressor-operator complexes by the Escherichia coli integration host factor protein. Mol Microbiol. 1992 Jun;6(12):1715–1722. doi: 10.1111/j.1365-2958.1992.tb00896.x. [DOI] [PubMed] [Google Scholar]
  20. Gille H., Egan J. B., Roth A., Messer W. The FIS protein binds and bends the origin of chromosomal DNA replication, oriC, of Escherichia coli. Nucleic Acids Res. 1991 Aug 11;19(15):4167–4172. doi: 10.1093/nar/19.15.4167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Goosen N., van Heuvel M., Moolenaar G. F., van de Putte P. Regulation of Mu transposition. II. The escherichia coli HimD protein positively controls two repressor promoters and the early promoter of bacteriophage Mu. Gene. 1984 Dec;32(3):419–426. doi: 10.1016/0378-1119(84)90017-9. [DOI] [PubMed] [Google Scholar]
  22. Groenen M. A., Timmers E., van de Putte P. DNA sequences at the ends of the genome of bacteriophage Mu essential for transposition. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2087–2091. doi: 10.1073/pnas.82.7.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Higgins N. P., Collier D. A., Kilpatrick M. W., Krause H. M. Supercoiling and integration host factor change the DNA conformation and alter the flow of convergent transcription in phage Mu. J Biol Chem. 1989 Feb 15;264(5):3035–3042. [PubMed] [Google Scholar]
  24. Higgins N. P. Death and transfiguration among bacteria. Trends Biochem Sci. 1992 Jun;17(6):207–211. doi: 10.1016/0968-0004(92)90376-k. [DOI] [PubMed] [Google Scholar]
  25. Hodges-Garcia Y., Hagerman P. J., Pettijohn D. E. DNA ring closure mediated by protein HU. J Biol Chem. 1989 Sep 5;264(25):14621–14623. [PubMed] [Google Scholar]
  26. Howe M. M. Prophage deletion mapping of bacteriophage Mu-1. Virology. 1973 Jul;54(1):93–101. doi: 10.1016/0042-6822(73)90118-9. [DOI] [PubMed] [Google Scholar]
  27. Huisman O., Faelen M., Girard D., Jaffé A., Toussaint A., Rouvière-Yaniv J. Multiple defects in Escherichia coli mutants lacking HU protein. J Bacteriol. 1989 Jul;171(7):3704–3712. doi: 10.1128/jb.171.7.3704-3712.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hübner P., Arber W. Mutational analysis of a prokaryotic recombinational enhancer element with two functions. EMBO J. 1989 Feb;8(2):577–585. doi: 10.1002/j.1460-2075.1989.tb03412.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Johnson R. C., Bruist M. F., Simon M. I. Host protein requirements for in vitro site-specific DNA inversion. Cell. 1986 Aug 15;46(4):531–539. doi: 10.1016/0092-8674(86)90878-0. [DOI] [PubMed] [Google Scholar]
  30. Kahmann R. Methylation regulates the expression of a DNA-modification function encoded by bacteriophage Mu. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):639–646. doi: 10.1101/sqb.1983.047.01.075. [DOI] [PubMed] [Google Scholar]
  31. Kamp D., Kahmann R. Two pathways in bacteriophage Mu transposition? Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):329–336. doi: 10.1101/sqb.1981.045.01.046. [DOI] [PubMed] [Google Scholar]
  32. Kano Y., Goshima N., Wada M., Imamoto F. Participation of hup gene product in replicative transposition of Mu phage in Escherichia coli. Gene. 1989;76(2):353–358. doi: 10.1016/0378-1119(89)90175-3. [DOI] [PubMed] [Google Scholar]
  33. Koch C., Kahmann R. Purification and properties of the Escherichia coli host factor required for inversion of the G segment in bacteriophage Mu. J Biol Chem. 1986 Nov 25;261(33):15673–15678. [PubMed] [Google Scholar]
  34. Koch C., Vandekerckhove J., Kahmann R. Escherichia coli host factor for site-specific DNA inversion: cloning and characterization of the fis gene. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4237–4241. doi: 10.1073/pnas.85.12.4237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Krause H. M., Higgins N. P. On the mu repressor and early DNA intermediates of transposition. Cold Spring Harb Symp Quant Biol. 1984;49:827–834. doi: 10.1101/sqb.1984.049.01.093. [DOI] [PubMed] [Google Scholar]
  36. Krause H. M., Higgins N. P. Positive and negative regulation of the Mu operator by Mu repressor and Escherichia coli integration host factor. J Biol Chem. 1986 Mar 15;261(8):3744–3752. [PubMed] [Google Scholar]
  37. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  38. Lathe R., Kieny M. P., Skory S., Lecocq J. P. Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini. DNA. 1984;3(2):173–182. doi: 10.1089/dna.1984.3.173. [DOI] [PubMed] [Google Scholar]
  39. Leach D., Symonds N. The isolation and characterisation of a plaque-forming derivative of bacteriophage Mu carrying a fragment of Tn3 conferring ampicillin resistance. Mol Gen Genet. 1979 May 4;172(2):179–184. doi: 10.1007/BF00268280. [DOI] [PubMed] [Google Scholar]
  40. Leung P. C., Teplow D. B., Harshey R. M. Interaction of distinct domains in Mu transposase with Mu DNA ends and an internal transpositional enhancer. Nature. 1989 Apr 20;338(6217):656–658. doi: 10.1038/338656a0. [DOI] [PubMed] [Google Scholar]
  41. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  42. Mizuuchi M., Baker T. A., Mizuuchi K. Assembly of the active form of the transposase-Mu DNA complex: a critical control point in Mu transposition. Cell. 1992 Jul 24;70(2):303–311. doi: 10.1016/0092-8674(92)90104-k. [DOI] [PubMed] [Google Scholar]
  43. Mizuuchi M., Mizuuchi K. Efficient Mu transposition requires interaction of transposase with a DNA sequence at the Mu operator: implications for regulation. Cell. 1989 Jul 28;58(2):399–408. doi: 10.1016/0092-8674(89)90854-4. [DOI] [PubMed] [Google Scholar]
  44. Mott J. E., Galloway J. L., Platt T. Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination. EMBO J. 1985 Jul;4(7):1887–1891. doi: 10.1002/j.1460-2075.1985.tb03865.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Nilsson L., Vanet A., Vijgenboom E., Bosch L. The role of FIS in trans activation of stable RNA operons of E. coli. EMBO J. 1990 Mar;9(3):727–734. doi: 10.1002/j.1460-2075.1990.tb08166.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Nilsson L., Verbeek H., Vijgenboom E., van Drunen C., Vanet A., Bosch L. FIS-dependent trans activation of stable RNA operons of Escherichia coli under various growth conditions. J Bacteriol. 1992 Feb;174(3):921–929. doi: 10.1128/jb.174.3.921-929.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ninnemann O., Koch C., Kahmann R. The E.coli fis promoter is subject to stringent control and autoregulation. EMBO J. 1992 Mar;11(3):1075–1083. doi: 10.1002/j.1460-2075.1992.tb05146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Polard P., Prère M. F., Chandler M., Fayet O. Programmed translational frameshifting and initiation at an AUU codon in gene expression of bacterial insertion sequence IS911. J Mol Biol. 1991 Dec 5;222(3):465–477. doi: 10.1016/0022-2836(91)90490-w. [DOI] [PubMed] [Google Scholar]
  49. Ross W., Thompson J. F., Newlands J. T., Gourse R. L. E.coli Fis protein activates ribosomal RNA transcription in vitro and in vivo. EMBO J. 1990 Nov;9(11):3733–3742. doi: 10.1002/j.1460-2075.1990.tb07586.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Schaeffer F., Kolb A., Buc H. Point mutations change the thermal denaturation profile of a short DNA fragment containing the lactose control elements. Comparison between experiment and theory. EMBO J. 1982;1(1):99–105. doi: 10.1002/j.1460-2075.1982.tb01131.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Shapiro J. A., Higgins N. P. Differential activity of a transposable element in Escherichia coli colonies. J Bacteriol. 1989 Nov;171(11):5975–5986. doi: 10.1128/jb.171.11.5975-5986.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  53. Stark M. J. Multicopy expression vectors carrying the lac repressor gene for regulated high-level expression of genes in Escherichia coli. Gene. 1987;51(2-3):255–267. doi: 10.1016/0378-1119(87)90314-3. [DOI] [PubMed] [Google Scholar]
  54. Summers W. C. A simple method for extraction of RNA from E. coli utilizing diethyl pyrocarbonate. Anal Biochem. 1970 Feb;33(2):459–463. doi: 10.1016/0003-2697(70)90316-7. [DOI] [PubMed] [Google Scholar]
  55. Surette M. G., Chaconas G. A protein factor which reduces the negative supercoiling requirement in the Mu DNA strand transfer reaction is Escherichia coli integration host factor. J Biol Chem. 1989 Feb 15;264(5):3028–3034. [PubMed] [Google Scholar]
  56. Surette M. G., Chaconas G. The Mu transpositional enhancer can function in trans: requirement of the enhancer for synapsis but not strand cleavage. Cell. 1992 Mar 20;68(6):1101–1108. doi: 10.1016/0092-8674(92)90081-m. [DOI] [PubMed] [Google Scholar]
  57. Surette M. G., Lavoie B. D., Chaconas G. Action at a distance in Mu DNA transposition: an enhancer-like element is the site of action of supercoiling relief activity by integration host factor (IHF). EMBO J. 1989 Nov;8(11):3483–3489. doi: 10.1002/j.1460-2075.1989.tb08513.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Thompson J. F., Landy A. Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. Nucleic Acids Res. 1988 Oct 25;16(20):9687–9705. doi: 10.1093/nar/16.20.9687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Thompson J. F., Moitoso de Vargas L., Koch C., Kahmann R., Landy A. Cellular factors couple recombination with growth phase: characterization of a new component in the lambda site-specific recombination pathway. Cell. 1987 Sep 11;50(6):901–908. doi: 10.1016/0092-8674(87)90516-2. [DOI] [PubMed] [Google Scholar]
  60. Toussaint A., Expert D., Desmet L. Simultaneous expression of a bacteriophage Mu transposase and repressor: a way of preventing killing due to mini-Mu replication. Mol Microbiol. 1991 Aug;5(8):2011–2019. doi: 10.1111/j.1365-2958.1991.tb00823.x. [DOI] [PubMed] [Google Scholar]
  61. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  62. Van Leerdam E., Karreman C., van de Putte P. Ner, a cro-like function of bacteriophage Mu. Virology. 1982 Nov;123(1):19–28. doi: 10.1016/0042-6822(82)90291-4. [DOI] [PubMed] [Google Scholar]
  63. Vogel J. L., Li Z. J., Howe M. M., Toussaint A., Higgins N. P. Temperature-sensitive mutations in the bacteriophage Mu c repressor locate a 63-amino-acid DNA-binding domain. J Bacteriol. 1991 Oct;173(20):6568–6577. doi: 10.1128/jb.173.20.6568-6577.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Zerbib D., Prentki P., Gamas P., Freund E., Galas D. J., Chandler M. Functional organization of the ends of IS1: specific binding site for an IS 1-encoded protein. Mol Microbiol. 1990 Sep;4(9):1477–1486. [PubMed] [Google Scholar]
  65. van Rijn P. A., van de Putte P., Goosen N. Analysis of the IHF binding site in the regulatory region of bacteriophage Mu. Nucleic Acids Res. 1991 Jun 11;19(11):2825–2834. doi: 10.1093/nar/19.11.2825. [DOI] [PMC free article] [PubMed] [Google Scholar]

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