1887

Abstract

DnaB and DnaI proteins conserved in low-GC content Gram-positive bacteria are apparently involved in helicase loading at the replication initiation site and during the restarting of stalled replication forks. In this study, we found five novel mutants and three novel mutants by screening 750 temperature-sensitive Gram-positive mutants. All of the mutants had a single amino acid substitution in either DnaB or DnaI that controlled temperature-sensitive growth, as confirmed by transduction experiments using phage 80. DNA synthesis as measured by [H]thymine incorporation, origin-to-terminus ratios and flow cytometric analysis revealed that the and mutants were unable to initiate DNA replication at restrictive temperatures, which is similar to previous findings in . Furthermore, some of the mutants were found to exhibit asynchrony in the initiation of DNA replication. Also, a fraction of the mutant cells showed arrested replication, and the mutant tested was sensitive to mitomycin C, which causes DNA lesions. These results suggest that DnaB and DnaI are required not only for replication initiation and but also for regulation of its synchrony, and they provide support for the involvement of DnaI activity in the restart of arrested replication forks .

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2007-10-01
2019-11-17
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