Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1978 Sep;27(3):791–799. doi: 10.1128/jvi.27.3.791-799.1978

Gene Expression and Stability of mRNA Affected by DNA-Arrested Synthesis in Gene 59, 46, and 47 Mutants of Bacteriophage T4

Jen-Leih Wu 1,†, Yun-Chi Yeh 1
PMCID: PMC525867  PMID: 702642

Abstract

The effect of bacteriophage T4 gene 59 mutations (DNA-arrested synthesis) on kinetics of DNA synthesis, gene expression, and stability of mRNA has been studied. When Escherichia coli B was infected by a T4 gene 59 mutant, DNA synthesis proceeded to increase linearly after initiation, but started to decrease at 8 min and was completely arrested at 12 min at 37°C. At various incubation temperatures (20 to 42°C), the initial rates and times of arrest of DNA synthesis were different, but the total amount of DNA synthesized was constant. This result supports the hypothesis that function of gene 59 is required for the conversion of 63S DNA molecules to other replicative intermediates (39). The abnormality in protein synthesis caused by gene 59 mutation is manifested by (i) a delayed shutoff in the expression of early proteins (gene 43, 46, 39, 52, 63, 42-45, and some unidentified proteins), (ii) a reduced rate of late gene expression (gene 34, 37, 18, 20, 23, wac, 24, 22, 38, and 19), and (iii) an absence of cleavage of certain late proteins (23, 24, IPIII and 22 to 23*, 24*, IPIII*, and small fragments). It appears that there was no effect on the expression of gene 33, 55, and 32 by a mutation in gene 59. Results obtained from an addition of rifampin at the prereplicative cycle after infection indicated that mRNA from genes 43, rIIA, 46, 39, 52, and 63 are more stable in T4amC5 (gene 59) than in wild-type-infected cells. mRNA remained functional longer in mutant-infected cells, and this may explain the prolonged synthesis of certain early proteins. The gene expression of other DNA arrested mutants—those in genes 46 and 47—showed a pattern of abnormal protein synthesis similar to that found in gene 59 mutant-infected cells, except more late proteins are synthesized. The gene expression in terms of phage DNA structure is discussed.

Full text

PDF
791

Images in this article

794Image
on p.794
796Image
on p.796

Selected References

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

  1. Alberts B. M., Frey L. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA. Nature. 1970 Sep 26;227(5265):1313–1318. doi: 10.1038/2271313a0. [DOI] [PubMed] [Google Scholar]
  2. Bachrach U., Benchetrit L. Studies on phage internal proteins. II. Cleavage of a precursor of internal proteins during the morphogenesis of bacteriophage T4. Virology. 1974 May;59(1):51–58. doi: 10.1016/0042-6822(74)90205-0. [DOI] [PubMed] [Google Scholar]
  3. Bolle A., Epstein R. H., Salser W., Geiduschek E. P. Transcription during bacteriophage T4 development: requirements for late messenger synthesis. J Mol Biol. 1968 Apr 28;33(2):339–362. doi: 10.1016/0022-2836(68)90193-9. [DOI] [PubMed] [Google Scholar]
  4. Bolund C. Influence of gene 55 on the regulation of synthesis of some early enzymes in bacteriophage T4-infected escherichia coli. J Virol. 1973 Jul;12(1):49–57. doi: 10.1128/jvi.12.1.49-57.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolund C., Sköld O. Regulation of early mRNA synthesis in bacteriophage T4-infected bacteria: dependence on bacteriophage-specific protein synthesis. J Virol. 1973 Jul;12(1):39–48. doi: 10.1128/jvi.12.1.39-48.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bruner R., Cape R. E. The expression of two classes of late genes of bacteriophage T4. J Mol Biol. 1970 Oct 14;53(1):69–89. doi: 10.1016/0022-2836(70)90046-x. [DOI] [PubMed] [Google Scholar]
  7. Cascino A., Geiduschek E. P., Cafferata R. L., Haselkorn R. T4 DNA replication and viral gene expression. J Mol Biol. 1971 Oct 28;61(2):357–367. doi: 10.1016/0022-2836(71)90385-8. [DOI] [PubMed] [Google Scholar]
  8. Guha A., Szybalski W., Salser W., Geiduschek E. P., Pulitzer J. F., Bolle A. Controls and polarity of transcription during bacteriophage T4 development. J Mol Biol. 1971 Jul 28;59(2):329–349. doi: 10.1016/0022-2836(71)90054-4. [DOI] [PubMed] [Google Scholar]
  9. Heil A., Zillig W. Reconstitution of bacterial DNA-dependent RNA-polymerase from isolated subunits as a tool for the elucidation of the role of the subunits in transcription. FEBS Lett. 1970 Dec;11(3):165–168. doi: 10.1016/0014-5793(70)80519-1. [DOI] [PubMed] [Google Scholar]
  10. Hosoda J., Cone R. Analysis of T4 phage proteins. I. Conversion of precursor proteins into lower molecular weight peptides during normal capsid formation. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1275–1281. doi: 10.1073/pnas.66.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KELLENBERGER E., SECHAUD J. Electron microscopical studies of phage multiplication. II. Production of phage-related structures during multiplication of phages T2 and T4. Virology. 1957 Apr;3(2):256–274. doi: 10.1016/0042-6822(57)90092-2. [DOI] [PubMed] [Google Scholar]
  12. Karam J. D., Bowles M. G. Mutation to overproduction of bacteriophage T4 gene products. J Virol. 1974 Feb;13(2):428–438. doi: 10.1128/jvi.13.2.428-438.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Karam J., McCulley C., Leach M. Genetic control of mRNA decay in T4 phage-infected Escherichia coli. Virology. 1977 Feb;76(2):685–700. doi: 10.1016/0042-6822(77)90251-3. [DOI] [PubMed] [Google Scholar]
  14. Kim J. S., Davidson N. Electron microscope heteroduplex study of sequence relations of T2, T4, and T6 bacteriophage DNAs. Virology. 1974 Jan;57(1):93–111. doi: 10.1016/0042-6822(74)90111-1. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  17. Linder C. H., Sköld O. Evidence for a diffusible T4 bacteriophage protein governing the initiation of delayed early RNA synthesis. J Virol. 1977 Jan;21(1):7–15. doi: 10.1128/jvi.21.1.7-15.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. O'Farrell P. Z., Gold L. M. Bacteriophage T4 gene expression. Evidence for two classes of prereplicative cistrons. J Biol Chem. 1973 Aug 10;248(15):5502–5511. [PubMed] [Google Scholar]
  19. Onorato L., Showe M. K. Gene 21 protein-dependent proteolysis in vitro of purified gene 22 product of bacteriophage T4. J Mol Biol. 1975 Mar 5;92(3):395–412. doi: 10.1016/0022-2836(75)90288-0. [DOI] [PubMed] [Google Scholar]
  20. Poglazov B. F., Levshenko M. T. Bacteriophage T4D-induced proteinase. J Mol Biol. 1974 Apr 15;84(3):463–467. doi: 10.1016/0022-2836(74)90452-5. [DOI] [PubMed] [Google Scholar]
  21. Ratner D. Letter to the editor: Bacteriophage T4 transcriptional control gene 55 codes for a protein bound to Escherichia coli RNA polymerase. J Mol Biol. 1974 Nov 15;89(4):803–807. doi: 10.1016/0022-2836(74)90054-0. [DOI] [PubMed] [Google Scholar]
  22. Riva S., Cascino A., Geiduschek E. P. Coupling of late transcription to viral replication in bacteriophage T4 development. J Mol Biol. 1970 Nov 28;54(1):85–102. doi: 10.1016/0022-2836(70)90447-x. [DOI] [PubMed] [Google Scholar]
  23. Riva S., Cascino A., Geiduschek E. P. Uncoupling of late transcription from DNA replication in bacteriophage T4 development. J Mol Biol. 1970 Nov 28;54(1):103–119. doi: 10.1016/0022-2836(70)90448-1. [DOI] [PubMed] [Google Scholar]
  24. Shah D. B., Berger H. Replication of gene 46-47 amber mutants of bacteriophage T4D. J Mol Biol. 1971 Apr 14;57(1):17–34. doi: 10.1016/0022-2836(71)90117-3. [DOI] [PubMed] [Google Scholar]
  25. Stevens A. New small polypeptides associated with DNA-dependent RNA polymerase of Escherichia coli after infection with bacteriophage T4. Proc Natl Acad Sci U S A. 1972 Mar;69(3):603–607. doi: 10.1073/pnas.69.3.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
  27. Taylor A. L., Trotter C. D. Linkage map of Escherichia coli strain K-12. Bacteriol Rev. 1972 Dec;36(4):504–524. doi: 10.1128/br.36.4.504-524.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vanderslice R. W., Yegian C. D. The identification of late bacteriophage T4 proteins on sodium dodecyl sulfate polyacrylamide gels. Virology. 1974 Jul;60(1):265–275. doi: 10.1016/0042-6822(74)90384-5. [DOI] [PubMed] [Google Scholar]
  29. WIBERG J. S., DIRKSEN M. L., EPSTEIN R. H., LURIA S. E., BUCHANAN J. M. Early enzyme synthesis and its control in E. coli infected with some amber mutants of bacteriophage T4. Proc Natl Acad Sci U S A. 1962 Feb;48:293–302. doi: 10.1073/pnas.48.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wiberg J. S., Mendelsohn S. L., Warner V., Aldrich C., Cardillo T. S. Genetic mapping of regA mutants of bacteriophage T4D. J Virol. 1977 Jun;22(3):742–749. doi: 10.1128/jvi.22.3.742-749.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wiberg J. S., Mendelsohn S., Warner V., Hercules K., Aldrich C., Munro J. L. SP62, a viable mutant of bacteriophage T4D defective in regulation of phage enzyme synthesis. J Virol. 1973 Oct;12(4):775–792. doi: 10.1128/jvi.12.4.775-792.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wu J. R., Yeh Y. C. New Late Gene, dar, Involved in DNA Replication of Bacteriophage T4 I. Isolation, Characterization, and Genetic Location. J Virol. 1975 May;15(5):1096–1106. doi: 10.1128/jvi.15.5.1096-1106.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wu J. R., Yeh Y. C. New late gene, dar, involved in the replication of bacteriophage T4 DNA. II. Overproduction of DNA binding protein (gene 32 protein) and further characterization. J Virol. 1978 Jul;27(1):90–102. doi: 10.1128/jvi.27.1.90-102.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wu J. R., Yeh Y. C. Requirement of a functional gene 32 product of bacteriophage T4 in UV, repair. J Virol. 1973 Oct;12(4):758–765. doi: 10.1128/jvi.12.4.758-765.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wu R., Wu J. L., Yeh Y. C. Role of gene 59 of bacteriophage T4 in repair of UV-irradiated and alkylated DNA in vivo. J Virol. 1975 Jul;16(1):5–16. doi: 10.1128/jvi.16.1.5-16.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wu R., Yeh Y. C. DNA arrested mutants of gene 59 of bacteriophage T4. II. Replicative intermediates. Virology. 1974 May;59(1):108–122. doi: 10.1016/0042-6822(74)90209-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES