| Autor: |
Bruand, Claude, Velten, Marion, McGovern, Stephen, Marsin, Stéphanie, Sérèna, Céline, Ehrlich, S. Dusko, Polard, Patrice |
| Předmět: |
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| Zdroj: |
Molecular Microbiology; Feb2005, Vol. 55 Issue 4, p1138-1150, 13p |
| Abstrakt: |
Initiation and re-initiation of chromosomal DNA replication in bacteria rely on divergent multiprotein assemblies, which direct the functional delivery of the replicative helicase on single-stranded DNA (ssDNA) at specific sites. These two processes are triggered either at the single chromosomal originoriCor at arrested forks by the conserved DnaA and PriA proteins respectively. InBacillus subtilis, these two pathways further require the three essential proteins DnaB, DnaD and DnaI, restrictively encoded in Gram positive bacteria of low GC content. We have recently shown that DnaI and DnaB act as a pair of loaders of the DnaC replicative helicase. The role of DnaD appeared more enigmatic. It was previously shown to interact with DnaA and to display weak ssDNA binding activity. Here, we report that purified DnaD can interact physically with PriA and with DnaB. We show that the lethality of the temperature-sensitivednaD23mutant can be suppressed by different DnaB point mutants, which were found to be identical to the suppressors ofpriAnull mutants. The DnaD23 protein displays lower ssDNA binding activity than DnaD. Conversely, the DnaB75 protein, the maindnaD23suppressor, has gained affinity for ssDNA. Finally, we observed that this interplay between DnaD and DnaB is crucial for their concerted interaction with SSB-coated ssDNA, which is the expected substrate for the loading of the replicative helicasein vivo. Altogether, these results highlight the need for both DnaD and DnaB to interact individually and together with ssDNA during the early stages of initiation and re-initiation of chromosomal DNA replication. They also point at a main structural role of DnaD in the multiprotein assemblies built during these two essential processes. [ABSTRACT FROM AUTHOR] |
| Databáze: |
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