Nucleic Acids Research, 1992, Vol. 20, No. 12 3029-3036
© 1992
MOLECULAR BIOLOGY |
Different effects of mioC transcription on initiation of chromosomal and minichromosomal replication in Escherichia coli
Department of Microbiology, The Technical University of Denmark Building 221, DK-2800 Lyngby, Copenhagen, Denmark 1Department of Biophysics, Institute for Cancer Research Montebello, 0310 Oslo, Norway
* To whom correspondence should be addressed
Received March 18, 1992. Revised May 18, 1992. Accepted May 18, 1992.
The mioC gene, which neighbors the chromosomal origin of replication (oriC) in Escherichia coli, has in a number of studies been implicated in the control of oriC initiation on minichromosomes. The present work reports on the construction of cells carrying different mioC mutations on the chromosome itself. Flow cytometry was employed to study the DNA replication control and growth pattern of the resulting mioC mutants. All parameters measured (growth rate, cell size, DNA/cell, number of origins per cell, timing of initiation) were the same for the wild type and all the mioC mutant cells under steady state growth and after different shifts in growth medium and after induction of the stringent response. It may be concluded that the dramatic effects of mioC mutations reported for minichromosomes are not observed for chromosomal replication and that the mioC gene and gene product is of little importance for the control of initiation. The data demonstrate that a minichromosome is not necessarily a valid model for chromosomal replication.
+ Present address: Department of Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309-0347, USA
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