Mechanism of Replication-Coupled DNA Interstrand Crosslink Repair

Markus Räschle, Puck Knipsheer, Milica Enoiu, Todor Angelov, Jingchuan Sun, Jack D. Griffith, Tom E. Ellenberger, Orlando D. Schärer, Johannes C. Walter

Research output: Contribution to journalArticle

321 Scopus citations

Abstract

DNA interstrand crosslinks (ICLs) are toxic DNA lesions whose repair occurs in the S phase of metazoans via an unknown mechanism. Here, we describe a cell-free system based on Xenopus egg extracts that supports ICL repair. During DNA replication of a plasmid containing a site-specific ICL, two replication forks converge on the crosslink. Subsequent lesion bypass involves advance of a nascent leading strand to within one nucleotide of the ICL, followed by incisions, translesion DNA synthesis, and extension of the nascent strand beyond the lesion. Immunodepletion experiments suggest that extension requires DNA polymerase ζ. Ultimately, a significant portion of the input DNA is fully repaired, but not if DNA replication is blocked. Our experiments establish a mechanism for ICL repair that reveals how this process is coupled to DNA replication.

Original languageEnglish
Pages (from-to)969-980
Number of pages12
JournalCell
Volume134
Issue number6
DOIs
StatePublished - Sep 19 2008

Keywords

  • DNA

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    Räschle, M., Knipsheer, P., Enoiu, M., Angelov, T., Sun, J., Griffith, J. D., Ellenberger, T. E., Schärer, O. D., & Walter, J. C. (2008). Mechanism of Replication-Coupled DNA Interstrand Crosslink Repair. Cell, 134(6), 969-980. https://doi.org/10.1016/j.cell.2008.08.030