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Frequent template switching in postreplication gaps: suppression of deleterious consequences by the Escherichia coli Uup and RadD proteins

Journal Article


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Abstract


  • © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. When replication forks encounter template DNA lesions, the lesion is simply skipped in some cases. The resulting lesion-containing gap must be converted to duplex DNA to permit repair. Some gap filling occurs via template switching, a process that generates recombination-like branched DNA intermediates. The Escherichia coli Uup and RadD proteins function in different pathways to process the branched intermediates. Uup is a UvrA-like ABC family ATPase. RadD is a RecQ-like SF2 family ATPase. Loss of both functions uncovers frequent and RecA-independent deletion events in a plasmid-based assay. Elevated levels of crossing over and repeat expansions accompany these deletion events, indicating that many, if not most, of these events are associated with template switching in postreplication gaps as opposed to simple replication slippage. The deletion data underpin simulations indicating that multiple postreplication gaps may be generated per replication cycle. Both Uup and RadD bind to branched DNAs in vitro. RadD protein suppresses crossovers and Uup prevents nucleoid mis-segregation. Loss of Uup and RadD function increases sensitivity to ciprofloxacin. We present Uup and RadD as genomic guardians. These proteins govern two pathways for resolution of branched DNA intermediates such that potentially deleterious genome rearrangements arising from frequent template switching are averted.

Authors


  •   Romero, Zachary (external author)
  •   Armstrong, Thomas (external author)
  •   Henrikus, Sarah (external author)
  •   Chen, Stefanie (external author)
  •   Glass, David (external author)
  •   Ferrazzoli, Alexander (external author)
  •   Wood, Elizabeth (external author)
  •   Chitteni-Pattu, Sindhu (external author)
  •   van Oijen, Antoine M.
  •   Lovett, Susan (external author)
  •   Robinson, Andrew
  •   Cox, Michael (external author)

Publication Date


  • 2020

Citation


  • Romero, Z., Armstrong, T., Henrikus, S., Chen, S., Glass, D., Ferrazzoli, A., Wood, E., Chitteni-Pattu, S., van Oijen, A., Lovett, S., Robinson, A. & Cox, M. (2020). Frequent template switching in postreplication gaps: suppression of deleterious consequences by the Escherichia coli Uup and RadD proteins. Nucleic acids research, 48 (1), 212-230.

Scopus Eid


  • 2-s2.0-85077477909

Ro Full-text Url


  • https://ro.uow.edu.au/cgi/viewcontent.cgi?article=2137&context=smhpapers1

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers1/1123

Number Of Pages


  • 18

Start Page


  • 212

End Page


  • 230

Volume


  • 48

Issue


  • 1

Place Of Publication


  • United Kingdom

Abstract


  • © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. When replication forks encounter template DNA lesions, the lesion is simply skipped in some cases. The resulting lesion-containing gap must be converted to duplex DNA to permit repair. Some gap filling occurs via template switching, a process that generates recombination-like branched DNA intermediates. The Escherichia coli Uup and RadD proteins function in different pathways to process the branched intermediates. Uup is a UvrA-like ABC family ATPase. RadD is a RecQ-like SF2 family ATPase. Loss of both functions uncovers frequent and RecA-independent deletion events in a plasmid-based assay. Elevated levels of crossing over and repeat expansions accompany these deletion events, indicating that many, if not most, of these events are associated with template switching in postreplication gaps as opposed to simple replication slippage. The deletion data underpin simulations indicating that multiple postreplication gaps may be generated per replication cycle. Both Uup and RadD bind to branched DNAs in vitro. RadD protein suppresses crossovers and Uup prevents nucleoid mis-segregation. Loss of Uup and RadD function increases sensitivity to ciprofloxacin. We present Uup and RadD as genomic guardians. These proteins govern two pathways for resolution of branched DNA intermediates such that potentially deleterious genome rearrangements arising from frequent template switching are averted.

Authors


  •   Romero, Zachary (external author)
  •   Armstrong, Thomas (external author)
  •   Henrikus, Sarah (external author)
  •   Chen, Stefanie (external author)
  •   Glass, David (external author)
  •   Ferrazzoli, Alexander (external author)
  •   Wood, Elizabeth (external author)
  •   Chitteni-Pattu, Sindhu (external author)
  •   van Oijen, Antoine M.
  •   Lovett, Susan (external author)
  •   Robinson, Andrew
  •   Cox, Michael (external author)

Publication Date


  • 2020

Citation


  • Romero, Z., Armstrong, T., Henrikus, S., Chen, S., Glass, D., Ferrazzoli, A., Wood, E., Chitteni-Pattu, S., van Oijen, A., Lovett, S., Robinson, A. & Cox, M. (2020). Frequent template switching in postreplication gaps: suppression of deleterious consequences by the Escherichia coli Uup and RadD proteins. Nucleic acids research, 48 (1), 212-230.

Scopus Eid


  • 2-s2.0-85077477909

Ro Full-text Url


  • https://ro.uow.edu.au/cgi/viewcontent.cgi?article=2137&context=smhpapers1

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers1/1123

Number Of Pages


  • 18

Start Page


  • 212

End Page


  • 230

Volume


  • 48

Issue


  • 1

Place Of Publication


  • United Kingdom