Inhibition of thymidine transport in dnaA mutants of Escherichia coli

Tohru Mizushima, Keiko Yokoyama, Shinji Mima, Tomofusa Tsuchiya, Kazuhisa Sekimizu

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17 Citations (Scopus)

Abstract

DnaA protein is the initiator of chromosomal DNA replication in Escherichia coli. We report here our evidence that thymidine transport across cytoplasmic membranes in temperature-sensitive dnaA mutants is greatly decreased at a permissive temperature of growth of the mutants. Complementation analysis with a plasmid containing the wild type dnaA gene and P1 phage-mediated transduction confirmed that mutations in the dnaA gene were responsible for the phenotype. A low level of nucleoside transport in the dnaA mutant was specific for thymidine; transport activities for other nucleosides were much the same as those in wild type cells. Membrane vesicles prepared from the dnaA mutant showed much the same activity of thymidine transport as did those from the wild type cells. No significant difference in the activity of thymidine kinase, which is known to facilitate thymidine transport, was seen between the mutant and the wild type cells. An increase in the pool of dTTP, a negative regulator for thymidine kinase, was observed in the dnaA mutant. Based on these observations, we suggest that inhibition of thymidine transport in dnaA mutants is caused by increases in the dTTP pool.

Original languageEnglish
Pages (from-to)21195-21200
Number of pages6
JournalJournal of Biological Chemistry
Volume272
Issue number34
DOIs
Publication statusPublished - 1997 Aug 22

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Mizushima, T., Yokoyama, K., Mima, S., Tsuchiya, T., & Sekimizu, K. (1997). Inhibition of thymidine transport in dnaA mutants of Escherichia coli. Journal of Biological Chemistry, 272(34), 21195-21200. https://doi.org/10.1074/jbc.272.34.21195