DnaA protein is sensitive to a soluble factor and is specifically inactivated for initiation of in vitro replication of the Escherichia coli minichromosome

T. Katayama, E. Crooke

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

DnaA protein loses the capacity to initiate chromosomal replication when treated with a soluble cell extract. This inactivation depends upon DNA and hydrolyzable ribonucleoside triphosphate. The extract does not affect the activities of other replicative proteins or the ability of DnaA to initiate replication of single-stranded DNA that contains a DnaA-binding hairpin, indicating that the inhibitory effect is specific for the action of DnaA at eriC. Gel filtration experiments implicate a 150-kDa factor as being responsible. Mutant DnaAcos protein, which causes over initiation in vivo, is insensitive to the inactivating factor, suggesting a requirement for this negative control in vivo. We propose that a soluble factor controls initiation through downregulation of DnaA protein.

Original languageEnglish
Pages (from-to)9265-9271
Number of pages7
JournalJournal of Biological Chemistry
Volume270
Issue number16
DOIs
Publication statusPublished - Jan 1 1995
Externally publishedYes

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Escherichia coli
Ribonucleosides
Peptide Initiation Factors
Proteins
Single-Stranded DNA
Mutant Proteins
Cell Extracts
Gel Chromatography
Down-Regulation
Gels
DNA
In Vitro Techniques
Experiments
triphosphoric acid

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "DnaA protein loses the capacity to initiate chromosomal replication when treated with a soluble cell extract. This inactivation depends upon DNA and hydrolyzable ribonucleoside triphosphate. The extract does not affect the activities of other replicative proteins or the ability of DnaA to initiate replication of single-stranded DNA that contains a DnaA-binding hairpin, indicating that the inhibitory effect is specific for the action of DnaA at eriC. Gel filtration experiments implicate a 150-kDa factor as being responsible. Mutant DnaAcos protein, which causes over initiation in vivo, is insensitive to the inactivating factor, suggesting a requirement for this negative control in vivo. We propose that a soluble factor controls initiation through downregulation of DnaA protein.",
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