Genetic identification of two distinct DNA polymerases, DnaE and PolC, that are essential for chromosomal dna replication in staphylococcus aureus

R. Inoue, C. Kaito, M. Tanabe, K. Kamura, N. Akimitsu, K. Sekimizu

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

We isolated and characterized temperature-sensitive mutants for two genes, dnaE and polC, that are essential for DNA replication in Staphylococcus aureus. DNA replication in these mutants had a slow-stop phenotype when the temperature was shifted to a nonpermissive level. The dnaE gene encodes a homolog of the α-subunit of the DNA polymerase III holoenzyme, the replicase essential for chromosomal DNA replication in Escherichia coli. The polC gene encodes PolC, another catalytic subunit of DNA polymerase, which is specifically round in gram-positive bacteria. The wildtype dnaE or polC gene complemented the temperature-sensitive phenotypes of cell growth and DNA replication in the corresponding mutant. Single mutations resulting in amino-acid exchanges were identified in the dnaE and polC genes of the temperature-sensitive mutants. The results indicate that these genes encode two distinct DNA polymerases which are both essential for chromosomal DNA replication in S. aureus. The number of viable mutant cells decreased at non-permissive temperature, suggesting that inactivation of DnaE and PolC has a bactericidal effect and that these enzymes are potential targets of antibiotics.

Original languageEnglish
Pages (from-to)564-571
Number of pages8
JournalMolecular Genetics and Genomics
Volume266
Issue number4
DOIs
Publication statusPublished - 2001
Externally publishedYes

Keywords

  • DNA polymerase
  • DNA replication
  • DnaE
  • PolC
  • Staphylococcus aureus

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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