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Single-strand DNA breaks cause replisome disassembly.

Kyle B Vrtis1, James M Dewar1, Gheorghe Chistol1

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA.

Molecular Cell
|January 23, 2021
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Summary
This summary is machine-generated.

DNA nicks, abundant in cells, unexpectedly cause replication fork collapse by creating double-strand breaks. The CMG helicase (CDC45-MCM2-7-GINS) disassembles upon encountering these breaks, highlighting nicks as uniquely dangerous DNA lesions.

Keywords:
CMGDNA repairDNA replicationdouble-strand breakfork collapsehomologous recombinationsingle moleculesingle-strand break

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA damage impedes replication and threatens genome stability.
  • The CMG helicase (CDC45-MCM2-7-GINS) stalls at DNA adducts but usually resumes replication.
  • The impact of single-strand breaks (nicks) on replication is poorly understood.

Purpose of the Study:

  • To investigate how the CMG helicase responds to DNA nicks during replication.
  • To determine the consequences of CMG encountering nicks on leading and lagging strands.

Main Methods:

  • Utilized Xenopus egg extracts for in vitro replication studies.
  • Examined CMG helicase behavior upon encountering DNA nicks.

Main Results:

  • CMG collision with a leading strand nick creates a double-strand break (DSB), causing CMG to run off the break.
  • CMG encountering a lagging strand nick leads to translocation beyond the break, ubiquitylation, and removal from chromatin.
  • Nicks invariably cause replisome disassembly, unlike other DNA adducts.

Conclusions:

  • DNA nicks are uniquely dangerous lesions that lead to replication fork collapse.
  • The CMG helicase is removed from chromatin upon encountering nicks, preventing its storage on double-stranded DNA during replication stress resolution.