Inhibition of thymidine transport in dnaA mutants of Escherichia coli

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

Research output: Contribution to journalArticle

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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
Externally publishedYes

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Thymidine
Escherichia coli
Thymidine Kinase
Nucleosides
Genes
Bacteriophage P1
Membranes
Bacteriophages
Temperature
DNA Replication
Plasmids
Cell Membrane
Phenotype
Mutation
DNA
Growth
Proteins
thymidine 5'-triphosphate

ASJC Scopus subject areas

  • Biochemistry

Cite this

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

Inhibition of thymidine transport in dnaA mutants of Escherichia coli. / Mizushima, Tohru; Yokoyama, Keiko; Mima, Shinji; Tsuchiya, Tomofusa; Sekimizu, Kazuhisa.

In: Journal of Biological Chemistry, Vol. 272, No. 34, 22.08.1997, p. 21195-21200.

Research output: Contribution to journalArticle

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, vol. 272, no. 34, pp. 21195-21200. https://doi.org/10.1074/jbc.272.34.21195
Mizushima T, Yokoyama K, Mima S, Tsuchiya T, Sekimizu K. Inhibition of thymidine transport in dnaA mutants of Escherichia coli. Journal of Biological Chemistry. 1997 Aug 22;272(34):21195-21200. https://doi.org/10.1074/jbc.272.34.21195
Mizushima, Tohru ; Yokoyama, Keiko ; Mima, Shinji ; Tsuchiya, Tomofusa ; Sekimizu, Kazuhisa. / Inhibition of thymidine transport in dnaA mutants of Escherichia coli. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 34. pp. 21195-21200.
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