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Vol. 12, No. 19, pp. 3032-3043, October 1, 1998
1 Division of Biological Science, Graduate School of
Science, Nagoya University Chikusa-ku, Nagoya 464-8602, Japan;
2 Department of Cell Biology, Graduate School of Biological
Sciences, Nara Institute of Science and Technology,
Takayama-cho, Ikoma 630-0101, Japan
Replication of the Escherichia coli chromosome is initiated
at a unique site, oriC. Concurrent initiation occurs at all
oriC sites present in a cell once, and only once, per cell
cycle. A mechanism to ensure cyclic initiation events was found
operating through the chromosomal site, datA, a 1-kb segment
located at 94.7 min on the genetic map that titrates exceptionally
large amounts of the bacterial initiator protein, DnaA. A strain
lacking datA grew normally but exhibited an asynchronous
initiation phenotype as a result of extra initiation events. This
mutant phenotype was suppressed by DnaA-titrating plasmids.
Furthermore, mutations in a 9-bp DnaA-binding sequence (the DnaA box)
in datA were enough to induce the mutant phenotype. Thus,
datA is a novel chromosomal element that appears to adjust a
balance between free and bound DnaA for a single initiation event at a
fixed time in the bacterial cell cycle. Titration of DnaA to newly
duplicated datA during oriC sequestration, which is
mediated by hemimethylated GATC sequences in oriC and the SeqA
protein, would contribute to prevention of reinitiations when
oriC is desequestered.
[Key Words: datA; DnaA; initiation; oriC; replication]
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