The Role of Escherichia coli dnaA Gene and Its Integrative Suppression in M 13 Coliphage DNA Synthesis

Abstract

An F+ derivative of E. coli E508 thermosensitive in dnaA function (involved in DNA synthesis initiation), its revertant and an Hfr derivative of E508 (ts) in which the temperature-sensitive phenotype is suppressed by integrative suppression were compared for their ability to support M13 phage DNA synthesis at the nonpermissive temperature. Upon infection at the nonpermissive temperature, the revertant and the Hfr strain support normal phage replication while the temperature-sensitive mutant does not. When infection is carried out at a permissive temperature and the temperature is shifted up after infection, phage synthesis occurs in the temperature-sensitive mutant also, but in lesser quantity than in the revertant strain. Analysis of intracellular labeled phage DNA indicates: parental replicative form DNA synthesis is not dependent on dnaA function; progeny replicative form DNA synthesis is strongly inhibited in the temperature-sensitive dnaA mutant at the nonpermissive temperature; progeny single-strand DNA synthesis does not absolutely require dnaA function; progeny single-strand DNA is present in the circular form. The implication of the host DNA replication in M13 DNA synthesis is discussed.