Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 Oct;6(10):3191–3199. doi: 10.1002/j.1460-2075.1987.tb02631.x

The cox protein of bacteriophage P2 inhibits the formation of the repressor protein and autoregulates the early operon.

S Saha 1, E Haggård-Ljungquist 1, K Nordström 1
PMCID: PMC553762  PMID: 2826134

Abstract

The cox gene is the first gene of the early operon of bacteriophage P2. The early promoter Pe and the repressor promoter Pc are located close to each other and in such a way that their transcripts have opposite polarity and show an overlap of about 30 nucleotides. The expression of the early operon and of the C gene was studied in vivo by using fusions to a promoterless cat (chloramphenicol acetyl transferase) gene. The results show that the Cox protein negatively autoregulates the early operon and inhibits the formation of the repressor C. By measuring the efficiency of plating of a series of P2 virulent deletion mutants on bacteria carrying a cloned cox gene, the site of action of the Cox protein was mapped within the Pe-Pc region. The stimulatory effect of the C protein on expression of the Pc promoter was found to be due to inhibition of transcription from Pc; this was demonstrated by mutating Pe which showed that loss of transcription from Pe stimulated transcription from Pc. Hence, this is a case of regulation of gene expression by convergent transcription. By cloning the region C-Pe-Pc-cox such that the cat and kan genes are expressed from Pc and Pe, respectively, it was shown that only one of the resistances (Cm and Km) was expressed. This mimics the choice between lysogeny and lytic growth of the phage. The 'lysogenic' state was very stable whereas the 'lytic' state flipped to the 'lysogenic' at a somewhat higher frequency. The presence of a cloned cox gene drastically stimulated the formation of free phage from a P2-lysogen and dramatically reduced the frequency of lysogenization after P2 infection. We conclude that the pleiotropic effects of the cox (control of excision) gene, namely effects on lysogenization, formation of free phage and site-specific P2 recombination, can be explained by the effect of the Cox protein on the activity of the promoters Pc and Pe.

Full text

PDF
3191

Images in this article

3193Fig. 2.
on p.3193

Selected References

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

  1. BERTANI G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol. 1951 Sep;62(3):293–300. doi: 10.1128/jb.62.3.293-300.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BERTANI L. E. The effect of the inhibition of protein synthesis on the establishment of lysogeny. Virology. 1957 Aug;4(1):53–71. doi: 10.1016/0042-6822(57)90043-0. [DOI] [PubMed] [Google Scholar]
  3. Bertani G., Ljungquist E., Jagusztyn-Krynicka K., Jupp S. Defective particle assembly in wild type P2 bacteriophage and its correction by the lg mutation. J Gen Virol. 1978 Feb;38(2):251–261. doi: 10.1099/0022-1317-38-2-251. [DOI] [PubMed] [Google Scholar]
  4. Bertani L. E. Abortive induction of bacteriophage P2. Virology. 1968 Sep;36(1):87–103. doi: 10.1016/0042-6822(68)90119-0. [DOI] [PubMed] [Google Scholar]
  5. Bertani L. E., Bertani G. Preparation and characterization of temperate, non-inducible bacteriophage P2 (host: Escherichia coli). J Gen Virol. 1970 Feb;6(2):201–212. doi: 10.1099/0022-1317-6-2-201. [DOI] [PubMed] [Google Scholar]
  6. Bertani L. E. Cold-sensitive mutations in the Z gene of prophage P2 that result in increased sensitivity of the lysogens to a low molecular weight product of the host bacteria. Mol Gen Genet. 1978 Oct 25;166(1):85–90. doi: 10.1007/BF00379732. [DOI] [PubMed] [Google Scholar]
  7. Bertani L. E. Genetic interaction between the nip1 mutation and genes affecting integration and excision in phage P2. Mol Gen Genet. 1980 Apr;178(1):91–99. doi: 10.1007/BF00267217. [DOI] [PubMed] [Google Scholar]
  8. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  9. Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. doi: 10.1016/0378-1119(84)90136-7. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Coleman J., Green P. J., Inouye M. The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes. Cell. 1984 Jun;37(2):429–436. doi: 10.1016/0092-8674(84)90373-8. [DOI] [PubMed] [Google Scholar]
  12. Geisselsoder J., Youdarian P., Dehò G., Chidambaram M., Goldstein R., Ljungquist E. Mutants of satellite virus P4 that cannot derepress their bacteriophage P2 helper. J Mol Biol. 1981 May 5;148(1):1–19. doi: 10.1016/0022-2836(81)90232-1. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Goosen N., van de Putte P. Role of ner protein in bacteriophage Mu transposition. J Bacteriol. 1986 Aug;167(2):503–507. doi: 10.1128/jb.167.2.503-507.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haggård-Ljungquist E., Kockum K., Bertani L. E. DNA sequences of bacteriophage P2 early genes cox and B and their regulatory sites. Mol Gen Genet. 1987 Jun;208(1-2):52–56. doi: 10.1007/BF00330421. [DOI] [PubMed] [Google Scholar]
  17. Izant J. G., Weintraub H. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell. 1984 Apr;36(4):1007–1015. doi: 10.1016/0092-8674(84)90050-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Light J., Molin S. Post-transcriptional control of expression of the repA gene of plasmid R1 mediated by a small RNA molecule. EMBO J. 1983;2(1):93–98. doi: 10.1002/j.1460-2075.1983.tb01387.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Light J., Molin S. The sites of action of the two copy number control functions of plasmid R1. Mol Gen Genet. 1982;187(3):486–493. doi: 10.1007/BF00332633. [DOI] [PubMed] [Google Scholar]
  21. Lin C. S. Nucleotide sequence of the essential region of bacteriophage P4. Nucleic Acids Res. 1984 Nov 26;12(22):8667–8684. doi: 10.1093/nar/12.22.8667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lindahl G., Hirota Y., Jacob F. On the process of cellular division in Escherichia coli: replication of the bacterial chromosome under control of prophage P2. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2407–2411. doi: 10.1073/pnas.68.10.2407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lindahl G., Sunshine M. Excision-deficient mutants of bacteriophage P2. Virology. 1972 Jul;49(1):180–187. doi: 10.1016/s0042-6822(72)80019-9. [DOI] [PubMed] [Google Scholar]
  24. Ljungquist E., Kockum K., Bertani L. E. DNA sequences of the repressor gene and operator region of bacteriophage P2. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3988–3992. doi: 10.1073/pnas.81.13.3988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Marinus M. G., Morris N. R. Biological function for 6-methyladenine residues in the DNA of Escherichia coli K12. J Mol Biol. 1974 May 15;85(2):309–322. doi: 10.1016/0022-2836(74)90366-0. [DOI] [PubMed] [Google Scholar]
  26. Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  28. Ptashne M., Jeffrey A., Johnson A. D., Maurer R., Meyer B. J., Pabo C. O., Roberts T. M., Sauer R. T. How the lambda repressor and cro work. Cell. 1980 Jan;19(1):1–11. doi: 10.1016/0092-8674(80)90383-9. [DOI] [PubMed] [Google Scholar]
  29. Saha S., Lundqvist B., Haggård-Ljungquist E. Autoregulation of bacteriophage P2 repressor. EMBO J. 1987 Mar;6(3):809–814. doi: 10.1002/j.1460-2075.1987.tb04823.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sasaki I., Bertani G. Growth abnormalities in Hfr derivatives of Escherichia coli strain C. J Gen Microbiol. 1965 Sep;40(3):365–376. doi: 10.1099/00221287-40-3-365. [DOI] [PubMed] [Google Scholar]
  31. Simons R. W., Kleckner N. Translational control of IS10 transposition. Cell. 1983 Sep;34(2):683–691. doi: 10.1016/0092-8674(83)90401-4. [DOI] [PubMed] [Google Scholar]
  32. Six E. W., Lindqvist B. H. Mutual derepression in the P2-P4 bacteriophage system. Virology. 1978 Jun 15;87(2):217–230. doi: 10.1016/0042-6822(78)90127-7. [DOI] [PubMed] [Google Scholar]
  33. Ward D. F., Murray N. E. Convergent transcription in bacteriophage lambda: interference with gene expression. J Mol Biol. 1979 Sep 15;133(2):249–266. doi: 10.1016/0022-2836(79)90533-3. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES