Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Sep;169(9):4076–4085. doi: 10.1128/jb.169.9.4076-4085.1987

Identification of polypeptides encoded by an Escherichia coli locus (hflA) that governs the lysis-lysogeny decision of bacteriophage lambda.

F Banuett, I Herskowitz
PMCID: PMC213711  PMID: 3040675

Abstract

We report the cloning of the Escherichia coli hflA locus, which governs stability of phage lambda cII protein and which has been proposed to encode or regulate a cII-specific protease. The hflA locus was cloned on an 18-kilobase DNA fragment by selecting for plasmids that carry the neighboring purA gene. The boundaries of hflA were delimited by analysis of deletions and insertions constructed in vitro and by use of transposon Tn1000. Maxicell analysis of the proteins encoded by the hflA-containing fragment shows that hflA consists of at least two nonoverlapping genes, hflC and hflK, encoding polypeptides of 37,000 (C) and 46,000 (K) daltons. We observe that insertions into one gene eliminate the corresponding polypeptide and greatly reduce synthesis of the other. We suggest that these two polypeptides (K and C) interact to form a multimeric complex and that free subunits are unstable. We have constructed two types of fusions between hflA and lacZ. One is an hflC-lacZ protein fusion constructed in vitro; the other is an hfl-lacZ operon fusion in which a Mu dX(Apr lac) has inserted into the hflK gene. We have used the operon fusion to infer the direction of transcription of the hflK gene--toward hflC and in the same direction as hflC. Last, we describe evidence that hflA contains an additional gene, hflX, encoding a 50,000-dalton polypeptide.

Full text

PDF
4076

Images in this article

4081Image
on p.4081
4081Image
on p.4081
4082Image
on p.4082
4083Image
on p.4083

Selected References

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

  1. Baker T. A., Howe M. M., Gross C. A. Mu dX, a derivative of Mu d1 (lac Apr) which makes stable lacZ fusions at high temperature. J Bacteriol. 1983 Nov;156(2):970–974. doi: 10.1128/jb.156.2.970-974.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banuett F., Hoyt M. A., McFarlane L., Echols H., Herskowitz I. hflB, a new Escherichia coli locus regulating lysogeny and the level of bacteriophage lambda cII protein. J Mol Biol. 1986 Jan 20;187(2):213–224. doi: 10.1016/0022-2836(86)90229-9. [DOI] [PubMed] [Google Scholar]
  3. Belfort M., Wulff D. L. An analysis of the processes of infection and induction of E. coli mutant hfl-1 by bacteriophage lambda. Virology. 1973 Sep;55(1):183–192. doi: 10.1016/s0042-6822(73)81020-7. [DOI] [PubMed] [Google Scholar]
  4. Belfort M., Wulff D. L. Genetic and biochemical investigation of the Escherichia coli mutant hfl-1 which is lysogenized at high frequency by bacteriophage lambda. J Bacteriol. 1973 Jul;115(1):299–306. doi: 10.1128/jb.115.1.299-306.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Belfort M., Wulff D. The roles of the lambda c3 gene and the Escherichia coli catabolite gene activation system in the establishment of lysogeny by bacteriophage lambda. Proc Natl Acad Sci U S A. 1974 Mar;71(3):779–782. doi: 10.1073/pnas.71.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casadaban M. J., Martinez-Arias A., Shapira S. K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast. Methods Enzymol. 1983;100:293–308. doi: 10.1016/0076-6879(83)00063-4. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Dietlein G., Schweizer E. Control of fatty-acid synthetase biosynthesis in Saccharomyces cerevisiae. Eur J Biochem. 1975 Oct 1;58(1):177–184. doi: 10.1111/j.1432-1033.1975.tb02362.x. [DOI] [PubMed] [Google Scholar]
  10. Elledge S. J., Walker G. C. Proteins required for ultraviolet light and chemical mutagenesis. Identification of the products of the umuC locus of Escherichia coli. J Mol Biol. 1983 Feb 25;164(2):175–192. doi: 10.1016/0022-2836(83)90074-8. [DOI] [PubMed] [Google Scholar]
  11. Gautsch J. W., Wulff D. L. Fine structure mapping, complementation, and physiology of Escherichia coli hfl mutants. Genetics. 1974 Jul;77(3):435–448. doi: 10.1093/genetics/77.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Herskowitz I., Hagen D. The lysis-lysogeny decision of phage lambda: explicit programming and responsiveness. Annu Rev Genet. 1980;14:399–445. doi: 10.1146/annurev.ge.14.120180.002151. [DOI] [PubMed] [Google Scholar]
  15. Herskowitz I., Signer E. R. A site essential for expression of all late genes in bacteriophage lambda. J Mol Biol. 1970 Feb 14;47(3):545–556. doi: 10.1016/0022-2836(70)90321-9. [DOI] [PubMed] [Google Scholar]
  16. Hoopes B. C., McClure W. R. A cII-dependent promoter is located within the Q gene of bacteriophage lambda. Proc Natl Acad Sci U S A. 1985 May;82(10):3134–3138. doi: 10.1073/pnas.82.10.3134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hoyt M. A., Knight D. M., Das A., Miller H. I., Echols H. Control of phage lambda development by stability and synthesis of cII protein: role of the viral cIII and host hflA, himA and himD genes. Cell. 1982 Dec;31(3 Pt 2):565–573. doi: 10.1016/0092-8674(82)90312-9. [DOI] [PubMed] [Google Scholar]
  18. Jackson C. M., Nemerson Y. Blood coagulation. Annu Rev Biochem. 1980;49:765–811. doi: 10.1146/annurev.bi.49.070180.004001. [DOI] [PubMed] [Google Scholar]
  19. Jones M. O., Herskowitz I. Mutants of bacteriophage lambda which do not requre the cIII gene for efficient lysogenization. Virology. 1978 Jul 15;88(2):199–212. doi: 10.1016/0042-6822(78)90277-5. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Ljungquist E., Bukhari A. I. State of prophage Mu DNA upon induction. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3143–3147. doi: 10.1073/pnas.74.8.3143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. O'Connor M. B., Malamy M. H. A new insertion sequence, IS121, is found on the Mu dI1 (Ap lac) bacteriophage and the Escherichia coli K-12 chromosome. J Bacteriol. 1983 Nov;156(2):669–679. doi: 10.1128/jb.156.2.669-679.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roberts G. P., Brill W. J. Genetics and regulation of nitrogen fixation. Annu Rev Microbiol. 1981;35:207–235. doi: 10.1146/annurev.mi.35.100181.001231. [DOI] [PubMed] [Google Scholar]
  24. Roberts G. P., MacNeil T., MacNeil D., Brill W. J. Regulation and characterization of protein products coded by the nif (nitrogen fixation) genes of Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):267–279. doi: 10.1128/jb.136.1.267-279.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Sancar A., Kacinski B. M., Mott D. L., Rupp W. D. Identification of the uvrC gene product. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5450–5454. doi: 10.1073/pnas.78.9.5450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. Schweizer E., Werkmeister K., Jain M. K. Fatty acid biosynthesis in yeast. Mol Cell Biochem. 1978 Nov 1;21(2):95–107. doi: 10.1007/BF00240280. [DOI] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Taylor D. P., Cohen S. N. Structural and functional analysis of cloned DNA segments containing the replication and incompatibility regions of a miniplasmid derived from a copy number mutant of NR1. J Bacteriol. 1979 Jan;137(1):92–104. doi: 10.1128/jb.137.1.92-104.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Timmis K. N., Cabello F., Cohen S. N. Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6. Mol Gen Genet. 1978 Jun 14;162(2):121–137. doi: 10.1007/BF00267869. [DOI] [PubMed] [Google Scholar]

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

RESOURCES