Correlation of gene transcription with the time of initiation of chromosome replication in Escherichia coli

Authors

  • Patrick W. Theisen,

    1. Department of Biological Sciences, Florida Institute of Technology, 150 West University Boulevard, Melbourne, Florida 32901, USA.
    Search for more papers by this author
  • Julia E. Grimwade,

    1. Department of Biological Sciences, Florida Institute of Technology, 150 West University Boulevard, Melbourne, Florida 32901, USA.
    Search for more papers by this author
  • Alan C. Leonard,

    1. Department of Biological Sciences, Florida Institute of Technology, 150 West University Boulevard, Melbourne, Florida 32901, USA.
    Search for more papers by this author
  • Joseph A. Bogan,

    1. Department of Biological Sciences, Florida Institute of Technology, 150 West University Boulevard, Melbourne, Florida 32901, USA.
    Search for more papers by this author
  • Charles E. Helmstetter

    Corresponding author
    1. Department of Biological Sciences, Florida Institute of Technology, 150 West University Boulevard, Melbourne, Florida 32901, USA.
    • *For correspondence. Tel. (407) 768 8000, ext. 8788; Fax (407) 952 1818.

    Search for more papers by this author

Summary

Transcriptional levels of the Escherichia coli mioC and gidA genes, which flank the chromosomal origin of replication (oriC) and the dnaA gene, were correlated with the time of initiation of chromosome replication. The transcripts were measured either in dnaC2(ts) mutants that had been aligned for initiation of chromosome replication by a temperature shift or in synchronous cultures of cells obtained using the baby machine technique. In both types of experiments, mioC transcription was inhibited prior to initiation of chromosome replication and resumed several minutes after initiation. Conversely, gidA and dnaA transcription were both inhibited after initiation of replication, coincident with the period of hemimethylation of oriC DNA. It is proposed that mioC transcription prevents initiation of chromosome replication, and must terminate before replication can begin. It is further proposed that the eclipse period between rounds of replication, i.e. the minimum interval between successive initiations, encompasses the time required to methylate GATC sequences in newly replicated oriC plus the time required to terminate mioC transcription. Conversely, the active transcription of gidA and dnaA prior to initiation is consistent with their positive effects on initiation, and their shutdown after initiation could serve to limit premature reinitiation.

Ancillary