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- W3129293593 abstract "SUMMARY DNA replication in all organisms must overcome nucleoprotein blocks to complete genome duplication. Accessory replicative helicases in Escherichia coli , Rep and UvrD, help replication machinery overcome blocks by removing incoming nucleoprotein complexes or aiding the re-initiation of replication. Mechanistic details of Rep function have emerged from recent live cell studies; however, the division of UvrD between its multiple activities in DNA repair and its role as an accessory helicase remains unclear in live cells. Here, by integrating super-resolved single-molecule fluorescence microscopy with biochemical analysis, we find that UvrD self-associates into a tetramer and, unlike Rep, is not recruited to a specific replisome protein despite being found at approximately 80% of replication forks. Instead, its recruitment to forks is likely mediated by the very high frequency of replication blocks due to DNA bound proteins, including RNA polymerase and DNA damage. Deleting rep and DNA repair factor genes mutS and uvrA , and inhibiting transcription through RNA polymerase mutation and antibiotic inhibition, indicates that the level of UvrD at the fork is dependent on its function. Our findings show that UvrD is recruited to sites of nucleoprotein blocks via distinctly different mechanisms to Rep and plays a multi-faceted role in ensuring successful DNA replication." @default.
- W3129293593 created "2021-03-01" @default.
- W3129293593 creator A5035067312 @default.
- W3129293593 creator A5035913037 @default.
- W3129293593 creator A5037565193 @default.
- W3129293593 creator A5074311677 @default.
- W3129293593 creator A5075094817 @default.
- W3129293593 creator A5080646179 @default.
- W3129293593 creator A5091779052 @default.
- W3129293593 date "2021-02-22" @default.
- W3129293593 modified "2023-10-11" @default.
- W3129293593 title "Tetrameric UvrD helicase is located at the<i>E. coli</i>replisome due to frequent replication blocks" @default.
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