DnaC Inactivation in Escherichia coli K-12 Induces the SOS Response and Expression of Nucleotide Biosynthesis Genes

Research output: Contribution to journalJournal articleResearchpeer-review

BACKGROUND: Initiation of chromosome replication in E. coli requires the DnaA and DnaC proteins and conditionally-lethal dnaA and dnaC mutants are often used to synchronize cell populations.

METHODOLOGY/PRINCIPAL FINDINGS: DNA microarrays were used to measure mRNA steady-state levels in initiation-deficient dnaA46 and dnaC2 bacteria at permissive and non-permissive temperatures and their expression profiles were compared to MG1655 wildtype cells. For both mutants there was altered expression of genes involved in nucleotide biosynthesis at the non-permissive temperature. Transcription of the dnaA and dnaC genes was increased at the non-permissive temperature in the respective mutant strains indicating auto-regulation of both genes. Induction of the SOS regulon was observed in dnaC2 cells at 38 degrees C and 42 degrees C. Flow cytometric analysis revealed that dnaC2 mutant cells at non-permissive temperature had completed the early stages of chromosome replication initiation.

CONCLUSION/SIGNIFICANCE: We suggest that in dnaC2 cells the SOS response is triggered by persistent open-complex formation at oriC and/or by arrested forks that require DnaC for replication restart.

Original languageEnglish
Article numbere2984
JournalP L o S One
Volume3
Issue number8
Number of pages8
ISSN1932-6203
DOIs
Publication statusPublished - 2008
Externally publishedYes

    Research areas

  • Bacterial Proteins/genetics, Chromosomes, Bacterial/genetics, DNA Replication/genetics, DNA-Binding Proteins/genetics, Escherichia coli K12/genetics, Escherichia coli Proteins/antagonists & inhibitors, Flow Cytometry, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Genes, Bacterial, Genes, Lethal, Heat-Shock Proteins/genetics, Nucleotides/biosynthesis, Oligonucleotide Array Sequence Analysis, RNA, Bacterial/genetics, RNA, Messenger/genetics, SOS Response (Genetics), Thermodynamics, Transcription, Genetic

ID: 200971647