Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jun;71(6):4502–4508. doi: 10.1128/jvi.71.6.4502-4508.1997

Derepression of prophage P2 by satellite phage P4: cloning of the P4 epsilon gene and identification of its product.

T Liu 1, S K Renberg 1, E Haggård-Ljungquist 1
PMCID: PMC191670  PMID: 9151842

Abstract

Escherichia coli phage P4 lacks all of the genetic information necessary for capsid, tail, and lysis functions. P4 is therefore dependent on a helper phage, such as P2, for lytic propagation. During P4 superinfection of a P2 lysogen, the P2 prophage is derepressed by the action of the P4-encoded epsilon gene. We have cloned the epsilon gene and identified the 10-kDa E protein. The epsilon gene product is the only P4 protein required to derepress prophage P2, which leads to in situ P2 DNA replication. A two-plasmid derepression assay system has been developed to examine the derepression activity of E. The reporter plasmid contains the two face-to-face promoters, Pe and Pc, involved in the lysis-lysogeny transcriptional switch of phage P2 and the immunity repressor C. The Pe promoter is coupled to a cat reporter gene. In the construct, the C repressor is transcribed from the Pc promoter and represses the Pe promoter, which mimics the in situ-repressed P2 prophage. The E protein is supplied in trans from a compatible plasmid in which the epsilon gene is under the control of the T7 promoter. We show here that in the two-plasmid assay system, induction of the E protein derepresses the Pe promoter. The ash9 mutation, which is located upstream of the epsilon gene, enhances the E-mediated derepression of the Pe promoter. The purified E protein shows no specific DNA binding activity, and the implications of this are discussed.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Agarwal M., Arthur M., Arbeit R. D., Goldstein R. Regulation of icosahedral virion capsid size by the in vivo activity of a cloned gene product. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2428–2432. doi: 10.1073/pnas.87.7.2428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alano P., Dehò G., Sironi G., Zangrossi S. Regulation of the plasmid state of the genetic element P4. Mol Gen Genet. 1986 Jun;203(3):445–450. doi: 10.1007/BF00422069. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Calendar R., Ljungquist E., Deho G., Usher D. C., Goldstein R., Youderian P., Sironi G., Six E. W. Lysogenization by satellite phage P4. Virology. 1981 Aug;113(1):20–38. doi: 10.1016/0042-6822(81)90133-1. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Christie G. E., Calendar R. Interactions between satellite bacteriophage P4 and its helpers. Annu Rev Genet. 1990;24:465–490. doi: 10.1146/annurev.ge.24.120190.002341. [DOI] [PubMed] [Google Scholar]
  11. Dehò G., Ghisotti D., Alano P., Zangrossi S., Borrello M. G., Sironi G. Plasmid mode of propagation of the genetic element P4. J Mol Biol. 1984 Sep 15;178(2):191–207. doi: 10.1016/0022-2836(84)90139-6. [DOI] [PubMed] [Google Scholar]
  12. Dehó G., Zangrossi S., Ghisotti D., Sironi G. Alternative promoters in the development of bacteriophage plasmid P4. J Virol. 1988 May;62(5):1697–1704. doi: 10.1128/jvi.62.5.1697-1704.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dehó G., Zangrossi S., Sabbattini P., Sironi G., Ghisotti D. Bacteriophage P4 immunity controlled by small RNAs via transcription termination. Mol Microbiol. 1992 Nov;6(22):3415–3425. doi: 10.1111/j.1365-2958.1992.tb02209.x. [DOI] [PubMed] [Google Scholar]
  14. Diana C., Dehò G., Geisselsoder J., Tinelli L., Goldstein R. Viral interference at the level of capsid size determination by satellite phage P4. J Mol Biol. 1978 Dec 15;126(3):433–445. doi: 10.1016/0022-2836(78)90050-5. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Ghisotti D., Briani F., Forti F., Piazza F., Polo S., Sabbattini P., Sturniolo T., Terzano S., Zangrossi S., Zappone M. Multiple regulatory mechanisms controlling phage-plasmid P4 propagation. FEMS Microbiol Rev. 1995 Aug;17(1-2):127–134. doi: 10.1111/j.1574-6976.1995.tb00194.x. [DOI] [PubMed] [Google Scholar]
  17. Ghisotti D., Chiaramonte R., Forti F., Zangrossi S., Sironi G., Dehò G. Genetic analysis of the immunity region of phage-plasmid P4. Mol Microbiol. 1992 Nov;6(22):3405–3413. doi: 10.1111/j.1365-2958.1992.tb02208.x. [DOI] [PubMed] [Google Scholar]
  18. Goldstein R., Sedivy J., Ljungquist E. Propagation of satellite phage P4 as a plasmid. Proc Natl Acad Sci U S A. 1982 Jan;79(2):515–519. doi: 10.1073/pnas.79.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Halling C., Calendar R. Bacteriophage P2 ogr and P4 delta genes act independently and are essential for P4 multiplication. J Bacteriol. 1990 Jul;172(7):3549–3558. doi: 10.1128/jb.172.7.3549-3558.1990. [DOI] [PMC free article] [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. Bacteriophage P2: replication of the chromosome requires a protein which acts only on the genome that coded for it. Virology. 1970 Oct;42(2):522–533. doi: 10.1016/0042-6822(70)90295-3. [DOI] [PubMed] [Google Scholar]
  23. Lindqvist B. H., Dehò G., Calendar R. Mechanisms of genome propagation and helper exploitation by satellite phage P4. Microbiol Rev. 1993 Sep;57(3):683–702. doi: 10.1128/mr.57.3.683-702.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Liu Y., Haggård-Ljungquist E. Studies of bacteriophage P2 DNA replication: localization of the cleavage site of the A protein. Nucleic Acids Res. 1994 Dec 11;22(24):5204–5210. doi: 10.1093/nar/22.24.5204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ljungquist E., Bertani L. E. Properties and products of the cloned int gene of bacteriophage P2. Mol Gen Genet. 1983;192(1-2):87–94. doi: 10.1007/BF00327651. [DOI] [PubMed] [Google Scholar]
  26. Nilssen O., Fossdal C. G., Johansen B. V., Lindqvist B. H. Bacteriophage P4 capsid-size determination and its relationship to P2 helper interference. Virology. 1996 May 15;219(2):443–452. doi: 10.1006/viro.1996.0270. [DOI] [PubMed] [Google Scholar]
  27. Polo S., Sturniolo T., Dehó G., Ghisotti D. Identification of a phage-coded DNA-binding protein that regulates transcription from late promoters in bacteriophage P4. J Mol Biol. 1996 Apr 12;257(4):745–755. doi: 10.1006/jmbi.1996.0199. [DOI] [PubMed] [Google Scholar]
  28. Sabbattini P., Forti F., Ghisotti D., Dehò G. Control of transcription termination by an RNA factor in bacteriophage P4 immunity: identification of the target sites. J Bacteriol. 1995 Mar;177(6):1425–1434. doi: 10.1128/jb.177.6.1425-1434.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Saha S., Haggård-Ljungquist E., Nordström K. Activation of prophage P4 by the P2 Cox protein and the sites of action of the Cox protein on the two phage genomes. Proc Natl Acad Sci U S A. 1989 Jun;86(11):3973–3977. doi: 10.1073/pnas.86.11.3973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saha S., Haggård-Ljungquist E., Nordström K. The cox protein of bacteriophage P2 inhibits the formation of the repressor protein and autoregulates the early operon. EMBO J. 1987 Oct;6(10):3191–3199. doi: 10.1002/j.1460-2075.1987.tb02631.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shore D., Dehò G., Tsipis J., Goldstein R. Determination of capsid size by satellite bacteriophage P4. Proc Natl Acad Sci U S A. 1978 Jan;75(1):400–404. doi: 10.1073/pnas.75.1.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Six E. W., Klug C. A. Bacteriophage P4: a satellite virus depending on a helper such as prophage P2. Virology. 1973 Feb;51(2):327–344. doi: 10.1016/0042-6822(73)90432-7. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Six E. W., Sunshine M. G., Williams J., Haggård-Ljungquist E., Lindqvist B. H. Morphopoietic switch mutations of bacteriophage P2. Virology. 1991 May;182(1):34–46. doi: 10.1016/0042-6822(91)90645-r. [DOI] [PubMed] [Google Scholar]
  37. Six E. W. The helper dependence of satellite bacteriophage P4: which gene functions of bacteriophage P2 are needed by P4? Virology. 1975 Sep;67(1):249–263. doi: 10.1016/0042-6822(75)90422-5. [DOI] [PubMed] [Google Scholar]
  38. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  39. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  40. Terzano S., Christian R., Espinoza F. H., Calendar R., Dehò G., Ghisotti D. A new gene of bacteriophage P4 that controls DNA replication. J Bacteriol. 1994 Oct;176(19):6059–6065. doi: 10.1128/jb.176.19.6059-6065.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES