1887

Abstract

SeqA protein, a main negative regulator of the replication initiation of the chromosome, also has several other functions which are still poorly understood. It was demonstrated previously that in mutants the copy number of another replicon, the plasmid, is decreased, and that the activity of the promoter (whose function is required for stimulation of ) is lower than that in the wild-type host. Here, SeqA-mediated regulation of phage and plasmid replicons was investigated in more detail. No significant influence of SeqA on -dependent DNA replication was observed, indicating that a direct regulation of DNA replication by this protein is unlikely. On the other hand, density-shift experiments, in which the fate of labelled DNA was monitored after phage infection of host cells, strongly suggested the early appearance of replication intermediates and preferential rolling-circle replication of phage DNA in mutants. The directionality of plasmid replication in such mutants was, however, only slightly affected. The stability of the heritable replication complex was decreased in the mutant relative to the wild-type host, but a stable fraction of the O protein was easily detectable, indicating that such a heritable complex can function in the mutant. To investigate the influence of gene function on heritable complex- and transcription-dependent DNA replication, the efficiency of plasmid replication in amino acid-starved mutants was measured. Under these conditions, dysfunction resulted in impairment of plasmid replication. These results indicate that unlike , SeqA modulates DNA replication indirectly, most probably by influencing the stability of the replication complex and the transcriptional activation of .

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2007-05-01
2019-11-14
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