Repair of a DNA-protein crosslink by replication-coupled proteolysis

Research output: Contribution to journalJournal articlepeer-review

DNA-protein crosslinks (DPCs) are caused by environmental, endogenous, and chemotherapeutic agents and pose a severe threat to genome stability. We use Xenopus egg extracts to recapitulate DPC repair in vitro and show that this process is coupled to DNA replication. A DPC on the leading strand template arrests the replisome by stalling the CMG helicase. The DPC is then degraded on DNA, yielding a peptide-DNA adduct that is bypassed by CMG. The leading strand subsequently resumes synthesis, stalls again at the adduct, and then progresses past the adduct using DNA polymerase ζ. A DPC on the lagging strand template only transiently stalls the replisome, but it too is degraded, allowing Okazaki fragment bypass. Our experiments describe a versatile, proteolysis-based mechanism of S phase DPC repair that avoids replication fork collapse.

Original languageEnglish
JournalCell
Volume159
Issue number2
Pages (from-to)346-57
Number of pages12
ISSN0092-8674
DOIs
Publication statusPublished - 9 Oct 2014
Externally publishedYes

    Research areas

  • Animals, Cell Extracts, DNA Adducts, DNA Repair, DNA Replication, DNA-Directed DNA Polymerase, Genomic Instability, Ovum, Xenopus, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.

ID: 176967569