Formation of the right before the left mature DNA end during packaging-cleavage of bacteriophage T7 DNA concatemers

Philip Serwer, Robert H. Watson, Shirley J. Hayes

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

During bacteriophage T7 morphogenesis in a T7-infected cell, mature length T7 DNA molecules join end-to-end to form concatemers that are subsequently both packaged in the T7 capsid and cut to mature size. In the present study, the kinetics of the appearance in vivo of the mature right and left T7 DNA ends have been analyzed. To perform this analysis, the intercalating dye proflavine is used to interrupt DNA packaging. When used at 0.5 to 8.0 μg/ml, proflavine progressively inhibits events in the T7 DNA packaging pathway, without either altering protein synthesis or degrading intracellular T7 DNA. Restriction endonuclease kinetic analysis reveals that proflavine (8 μg/ml) completely blocks formation of the mature T7 DNA left end, but only partially blocks formation of the mature T7 DNA right end. Both these and other observations are explained by the hypothesis that, in the T7 DNA packaging pathway, events occur in the following sequence: (1) formation of a mature right end; (2) packaging of at least some of the genome; (3) formation of the mature left end.

Original languageEnglish (US)
Pages (from-to)311-317
Number of pages7
JournalJournal of Molecular Biology
Volume226
Issue number2
DOIs
StatePublished - Jul 20 1992
Externally publishedYes

Keywords

  • DNA packaging
  • agarose gel electrophoresis
  • bacteriophage
  • intercalator
  • restriction endonuclease

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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