Suppressors of DnaAATP imposed overinitiation in Escherichia coli

Godefroid Charbon, Leise Riber, Malene Cohen, Ole Skovgaard, Kazuyuki Fujimitsu, Tsutomu Katayama, Anders Løbner-Olesen

Research output: Contribution to journalArticle

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Abstract

Chromosome replication in Escherichia coli is limited by the supply of DnaA associated with ATP. Cells deficient in RIDA (Regulatory Inactivation of DnaA) due to a deletion of the hda gene accumulate suppressor mutations (hsm) to counteract the overinitiation caused by an elevated DnaAATP level. Eight spontaneous hda suppressor mutations were identified by whole-genome sequencing, and three of these were analysed further. Two mutations (hsm-2 and hsm-4) mapped in the dnaA gene and led to a reduced ability to initiate replication from oriC. One mutation (hsm-1) mapped to the seqA promoter and increased the SeqA protein level in the cell. hsm-1 cells had prolonged origin sequestration, reduced DnaA protein level and reduced DnaA-Reactivating Sequence (DARS)-mediated rejuvenation of DnaAADP to DnaAATP, all of which could contribute to the suppression of RIDA deficiency. Despite of these defects hsm-1 cells were quite similar to wild type with respect to cell cycle parameters. We speculate that since SeqA binding sites might overlap with DnaA binding sites spread throughout the chromosome, excess SeqA could interfere with DnaA titration and thereby increase free DnaA level. Thus, in spite of reduction in total DnaA, the amount of DnaA molecules available for initiation may not be reduced.

Original languageEnglish
Pages (from-to)914-928
Number of pages15
JournalMolecular Microbiology
Volume79
Issue number4
DOIs
Publication statusPublished - Feb 1 2011

Fingerprint

Escherichia coli
Genetic Suppression
Chromosomes
Binding Sites
Rejuvenation
Mutation
Gene Deletion
Cell Cycle
Proteins
Adenosine Triphosphate
Genome
Genes

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Charbon, G., Riber, L., Cohen, M., Skovgaard, O., Fujimitsu, K., Katayama, T., & Løbner-Olesen, A. (2011). Suppressors of DnaAATP imposed overinitiation in Escherichia coli. Molecular Microbiology, 79(4), 914-928. https://doi.org/10.1111/j.1365-2958.2010.07493.x

Suppressors of DnaAATP imposed overinitiation in Escherichia coli. / Charbon, Godefroid; Riber, Leise; Cohen, Malene; Skovgaard, Ole; Fujimitsu, Kazuyuki; Katayama, Tsutomu; Løbner-Olesen, Anders.

In: Molecular Microbiology, Vol. 79, No. 4, 01.02.2011, p. 914-928.

Research output: Contribution to journalArticle

Charbon, G, Riber, L, Cohen, M, Skovgaard, O, Fujimitsu, K, Katayama, T & Løbner-Olesen, A 2011, 'Suppressors of DnaAATP imposed overinitiation in Escherichia coli', Molecular Microbiology, vol. 79, no. 4, pp. 914-928. https://doi.org/10.1111/j.1365-2958.2010.07493.x
Charbon, Godefroid ; Riber, Leise ; Cohen, Malene ; Skovgaard, Ole ; Fujimitsu, Kazuyuki ; Katayama, Tsutomu ; Løbner-Olesen, Anders. / Suppressors of DnaAATP imposed overinitiation in Escherichia coli. In: Molecular Microbiology. 2011 ; Vol. 79, No. 4. pp. 914-928.
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