The G157C mutation in the Escherichia coli sliding clamp specifically affects initiation of replication.

Abstract:

Escherichia coli cells with a point mutation in the dnaN gene causing the amino acid change Gly157 to Cys, were found to underinitiate replication and grow with a reduced origin and DNA concentration. The mutant beta clamp also caused excessive conversion of ATP-DnaA to ADP-DnaA. The DnaA protein was, however, not the element limiting initiation of replication. Overproduction of DnaA protein, which in wild-type cells leads to over-replication, had no effect in the dnaN(G157C) mutant. Origins already opened by DnaA seemed to remain open for a prolonged period, with a stage of initiation involving beta clamp loading, presumably limiting the initiation process. The existence of opened origins led to a moderate SOS response. Lagging strand synthesis, which also requires loading of the beta clamp, was apparently unaffected. The result indicates that some aspects of beta clamp activity are specific to the origin. It is possible that the origin specific activities of beta contribute to regulation of initiation frequency.

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