Mutants of bacteriophage T4 deficient in the ability to induce nuclear disruption: shutoff of host DNA and protein synthesis, gene dosage experiments, identification of a restrictive host, and possible biological significance

D. P. Snustad, C. J.H. Bursch, D. A. Parson, S. H. Hefeneider

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

The shutoff of host DNA synthesis is delayed until about 8 to 10 min after infection when E. coli B/5 cells are infected with bacteriophage T4 mutants deficient in the ability to induce nuclear disruption (ndd mutants). The host DNA synthesized after infection with ndd mutants is stable in the absence of T4 endonucleases II and IV, but is unstable in the presence of these nucleases. Host protein synthesis, as indicated by the inducibility of β galactosidase and sodium dodecyl sulfate polyacrylamide gel patterns of isotopically labeled proteins synthesized after infection, is shut off normally in ndd infected cells, even in the absence of host DNA degradation. The Cal Tech wild type strain of E. coli CT447 was found to restrict growth of the ndd mutants. Since T4D+ also has a very low efficiency of plating on CT447, the authors isolated a nitrosoguanidine induced derivative of CT447 which yields a high T4D+ efficiency of plating while still restricting the ndd mutants. Using this derivative, CT447 T4 plq+ (for T4 plaque+), the authors have shown that host DNA degradation and shutoff of host DNA synthesis occur after infection with either ndd98 x 5 (shutoff delayed) or T4D+ (shutoff normal) with approximately the same kinetics as in E. coli strain B/5. Nuclear disruption occurs after infection of CT447 with ndd+ phage, but not after infection with ndd- phage. The rate of DNA synthesis after infection of CT447 T4 plq+ with ndd98 x 5 is about 75% of the rate observed after infection with T4D+ while the burst size of ndd98 x 5 is only 3.5% of that of T4D+. The results of gene dosage experiments using the ndd restrictive host CT447 suggest that the ndd gene product is required in stoichiometric amounts. The observation by thin section electron microscopy of two distinct pools of DNA, one apparently phage DNA and the other host DNA, in cells infected with nuclear disruption proficient, host DNA degradation phage suggests that nuclear disruption may be a compartmentalization mechanism which separates the pathways of host DNA degradation and phage DNA biosynthesis.

Original languageEnglish (US)
Pages (from-to)268-288
Number of pages21
JournalJournal of virology
Volume18
Issue number1
DOIs
StatePublished - 1976

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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