Abstract
Temperate coliphage HK022 requires integration host factor (IHF) for lytic growth. The determinant responsible for this requirement was identified as a new gene (roi) located between genes P and Q. This gene encodes a DNA-binding protein (Roi) containing a helix-turn-helix motif. We have shown that Roi binds a site within its own gene that is closely linked to an IHF binding site. By gel retardation experiments, we have found that IHF binding stabilizes the interaction of Roi with its gene. We have isolated three independent phage mutants that are able to grow on an IHF- host. They carry different mutations scattered in the roi gene and specifying single amino-acid changes. The interactions of all three Roi mutant proteins with the Roi binding site differed from that of the wild type. Roi displays strong similarities, in its C-terminal half, to two putative DNA-binding proteins of bacteriophage P1: Ant1 and KilA. The mode of action of the Roi protein and the possibility that IHF is modulating the expression and/or the action of Roi are discussed.
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Selected References
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- 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]
- Atkinson B. L., Gottesman M. E. The Escherichia coli rpoB60 mutation blocks antitermination by coliphage HK022 Q-function. J Mol Biol. 1992 Sep 5;227(1):29–37. doi: 10.1016/0022-2836(92)90679-e. [DOI] [PubMed] [Google Scholar]
- Bednarz A. L., Boocock M. R., Sherratt D. J. Determinants of correct res site alignment in site-specific recombination by Tn3 resolvase. Genes Dev. 1990 Dec;4(12B):2366–2375. doi: 10.1101/gad.4.12b.2366. [DOI] [PubMed] [Google Scholar]
- Bujard H., Gentz R., Lanzer M., Stueber D., Mueller M., Ibrahimi I., Haeuptle M. T., Dobberstein B. A T5 promoter-based transcription-translation system for the analysis of proteins in vitro and in vivo. Methods Enzymol. 1987;155:416–433. doi: 10.1016/0076-6879(87)55028-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Campbell A. Comparative molecular biology of lambdoid phages. Annu Rev Microbiol. 1994;48:193–222. doi: 10.1146/annurev.mi.48.100194.001205. [DOI] [PubMed] [Google Scholar]
- Clerget M., Jin D. J., Weisberg R. A. A zinc-binding region in the beta' subunit of RNA polymerase is involved in antitermination of early transcription of phage HK022. J Mol Biol. 1995 May 12;248(4):768–780. doi: 10.1006/jmbi.1995.0259. [DOI] [PubMed] [Google Scholar]
- Ditto M. D., Roberts D., Weisberg R. A. Growth phase variation of integration host factor level in Escherichia coli. J Bacteriol. 1994 Jun;176(12):3738–3748. doi: 10.1128/jb.176.12.3738-3748.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Freundlich M., Ramani N., Mathew E., Sirko A., Tsui P. The role of integration host factor in gene expression in Escherichia coli. Mol Microbiol. 1992 Sep;6(18):2557–2563. doi: 10.1111/j.1365-2958.1992.tb01432.x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Gamas P., Chandler M. G., Prentki P., Galas D. J. Escherichia coli integration host factor binds specifically to the ends of the insertion sequence IS1 and to its major insertion hot-spot in pBR322. J Mol Biol. 1987 May 20;195(2):261–272. doi: 10.1016/0022-2836(87)90648-6. [DOI] [PubMed] [Google Scholar]
- Goosen N., van de Putte P. Regulation of Mu transposition. I. Localization of the presumed recognition sites for HimD and Ner functions controlling bacteriophage Mu transcription. Gene. 1984 Oct;30(1-3):41–46. doi: 10.1016/0378-1119(84)90103-3. [DOI] [PubMed] [Google Scholar]
- Griffo G., Oppenheim A. B., Gottesman M. E. Repression of the lambda pcin promoter by integrative host factor. J Mol Biol. 1989 Sep 5;209(1):55–64. doi: 10.1016/0022-2836(89)90169-1. [DOI] [PubMed] [Google Scholar]
- Hansen E. B. Structure and regulation of the lytic replicon of phage P1. J Mol Biol. 1989 May 5;207(1):135–149. doi: 10.1016/0022-2836(89)90445-2. [DOI] [PubMed] [Google Scholar]
- Heisig A., Riedel H. D., Dobrinski B., Lurz R., Schuster H. Organization of the immunity region immI of bacteriophage P1 and synthesis of the P1 antirepressor. J Mol Biol. 1989 Oct 20;209(4):525–538. doi: 10.1016/0022-2836(89)90591-3. [DOI] [PubMed] [Google Scholar]
- Henthorn K. S., Friedman D. I. Identification of related genes in phages phi 80 and P22 whose products are inhibitory for phage growth in Escherichia coli IHF mutants. J Bacteriol. 1995 Jun;177(11):3185–3190. doi: 10.1128/jb.177.11.3185-3190.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mahajna J., Oppenheim A. B., Rattray A., Gottesman M. Translation initiation of bacteriophage lambda gene cII requires integration host factor. J Bacteriol. 1986 Jan;165(1):167–174. doi: 10.1128/jb.165.1.167-174.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mozola M. A., Carver D. L., Friedman D. I. A phi 80 function inhibitory for growth of lambdoid phage in him mutants of Escherichia coli deficient in integration host factor. II. Physiological analysis of the abortive infection. Virology. 1985 Jan 30;140(2):328–341. doi: 10.1016/0042-6822(85)90369-1. [DOI] [PubMed] [Google Scholar]
- Mozola M. A., Friedman D. I. A phi 80 function inhibitory for growth of lambdoid phage in him mutants of Escherichia coli deficient in integration host factor. I. Genetic analysis of the Rha phenotype. Virology. 1985 Jan 30;140(2):313–327. doi: 10.1016/0042-6822(85)90368-x. [DOI] [PubMed] [Google Scholar]
- Nash H. A. Bending and supercoiling of DNA at the attachment site of bacteriophage lambda. Trends Biochem Sci. 1990 Jun;15(6):222–227. doi: 10.1016/0968-0004(90)90034-9. [DOI] [PubMed] [Google Scholar]
- Oberto J., Weisberg R. A., Gottesman M. E. Structure and function of the nun gene and the immunity region of the lambdoid phage HK022. J Mol Biol. 1989 Jun 20;207(4):675–693. doi: 10.1016/0022-2836(89)90237-4. [DOI] [PubMed] [Google Scholar]
- Retallack D. M., Johnson L. L., Ziegler S. F., Strauch M. A., Friedman D. I. A single-base-pair mutation changes the specificities of both a transcription activation protein and its binding site. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9562–9565. doi: 10.1073/pnas.90.20.9562. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sternberg N., Hoess R. The molecular genetics of bacteriophage P1. Annu Rev Genet. 1983;17:123–154. doi: 10.1146/annurev.ge.17.120183.001011. [DOI] [PubMed] [Google Scholar]
- Yang C. C., Nash H. A. The interaction of E. coli IHF protein with its specific binding sites. Cell. 1989 Jun 2;57(5):869–880. doi: 10.1016/0092-8674(89)90801-5. [DOI] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
