Abstract
The pif region of the F factor includes two genes, pifA and pifB, that lead to abortive T7 infection. We have identified a new gene in this region, pifC, by constructing an in vitro fusion of pif DNA at 41.6 kilobases on the F factor physical map to the lacZ gene. A PifC-LacZ fusion protein of 149,000 daltons has been identified by immunoprecipitation and polyacrylamide gel electrophoresis. This allows us to assign the N terminus of pifC to 42.5 kilobases on the F map. Using fusions of pifC, pifA, and pifB to lacZ, we have studied the regulation of pif gene expression and have shown that the product of pifC negatively controls its own expression and that of pifA and pifB.
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- Blumberg D. D., Mabie C. T., Malamy M. H. T7 protein synthesis in F-factor-containing cells: evidence for an episomally induced impairment of translation and relation to an alteration in membrane permeability. J Virol. 1975 Jan;17(1):94–105. doi: 10.1128/jvi.17.1.94-105.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukhari A. I., Zipser D. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments. Nat New Biol. 1973 Jun 20;243(129):238–241. doi: 10.1038/newbio243238a0. [DOI] [PubMed] [Google Scholar]
- Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang S., Cohen S. N. In vivo site-specific genetic recombination promoted by the EcoRI restriction endonuclease. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4811–4815. doi: 10.1073/pnas.74.11.4811. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dharmalingam K., Goldberg E. B. Restriction in vivo. III. General effects of glucosylation and restriction on phage T4 gene expression and replication. Virology. 1979 Jul 30;96(2):393–403. doi: 10.1016/0042-6822(79)90097-7. [DOI] [PubMed] [Google Scholar]
- Eichelaub R., Wehlmann H. Amber-mutants of plasmid mini-F defective in replication. Mol Gen Genet. 1980;180(1):201–204. doi: 10.1007/BF00267370. [DOI] [PubMed] [Google Scholar]
- Eichenlaub R., Figurski D., Helinski D. R. Bidirection replication from a unique origin in a mini-F plasmid. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1138–1141. doi: 10.1073/pnas.74.3.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guyer M. S., Figurski D., Davidson N. Electron microscope study of a plasmid chimera containing the replication region of the Escherichia coli F plasmid. J Bacteriol. 1976 Aug;127(2):988–997. doi: 10.1128/jb.127.2.988-997.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
- Kilbane J. J., Malamy M. H. F factor mobilization of non-conjugative chimeric plasmids in Escherichia coli: general mechanisms and a role for site-specific recA-independent recombination at orV1. J Mol Biol. 1980 Oct 15;143(1):73–93. doi: 10.1016/0022-2836(80)90125-4. [DOI] [PubMed] [Google Scholar]
- Komai N., Nishizawa T., Hayakawa Y., Murotsu T., Matsubara K. Detection and mapping of six miniF-encoded proteins by cloning analysis of dissected miniF segments. Mol Gen Genet. 1982;186(2):193–203. doi: 10.1007/BF00331850. [DOI] [PubMed] [Google Scholar]
- Lane D., Gardner R. C. Second EcoRI fragment of F capable of self-replication. J Bacteriol. 1979 Jul;139(1):141–151. doi: 10.1128/jb.139.1.141-151.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MAEKELAE O., MAEKELAE P. H., SOIKKELI S. SEX-SPECIFICITY OF THE BACTERIOPHAGE T7. Ann Med Exp Biol Fenn. 1964;42:188–195. [PubMed] [Google Scholar]
- Malamy M. H., Fiandt M., Szybalski W. Electron microscopy of polar insertions in the lac operon of Escherichia coli. Mol Gen Genet. 1972;119(3):207–222. doi: 10.1007/BF00333859. [DOI] [PubMed] [Google Scholar]
- Morrison T. G., Malamy M. H. Comparisons of F factors and R factors: existence of independent regulation groups in F factors. J Bacteriol. 1970 Jul;103(1):81–88. doi: 10.1128/jb.103.1.81-88.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morrison T. G., Malamy M. H. T7 translational control mechanisms and their inhibiton by F factors. Nat New Biol. 1971 May 12;231(19):37–41. doi: 10.1038/newbio231037a0. [DOI] [PubMed] [Google Scholar]
- Oliver D. B., Beckwith J. E. coli mutant pleiotropically defective in the export of secreted proteins. Cell. 1981 Sep;25(3):765–772. doi: 10.1016/0092-8674(81)90184-7. [DOI] [PubMed] [Google Scholar]
- Rotman G. S., Cooney R., Malamy M. H. Cloning of the pif region of the F sex factor and identification of a pif protein product. J Bacteriol. 1983 Jul;155(1):254–264. doi: 10.1128/jb.155.1.254-264.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sharp P. A., Hsu M. T., Otsubo E., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors. J Mol Biol. 1972 Nov 14;71(2):471–497. doi: 10.1016/0022-2836(72)90363-4. [DOI] [PubMed] [Google Scholar]
- Skurray R. A., Nagaishi H., Clark A. J. Molecular cloning of DNA from F sex factor of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1976 Jan;73(1):64–68. doi: 10.1073/pnas.73.1.64. [DOI] [PMC free article] [PubMed] [Google Scholar]