TY - JOUR
T1 - Mechanism of Replication-Coupled DNA Interstrand Crosslink Repair
AU - Räschle, Markus
AU - Knipsheer, Puck
AU - Enoiu, Milica
AU - Angelov, Todor
AU - Sun, Jingchuan
AU - Griffith, Jack D.
AU - Ellenberger, Tom E.
AU - Schärer, Orlando D.
AU - Walter, Johannes C.
N1 - Funding Information:
We are grateful to Alan D'Andrea for advice and encouragement during early stages of this project. This work was supported by NIH grants GM62267 (J.C.W.), GM55390 (T.E.E.), and GM31819 (J.D.G.), a Leukemia and Lymphoma Scholar Award (J.C.W.), a Dutch Cancer Society Fellowship (P.K.), and grants from the New York State Office of Science and Technology and Academic Research NYSTAR, C040069 (O.D.S.) and the Swiss Cancer League OCS-01413-080-2003 (O.D.S.). We thank Agata Smogorzewska, Ralph Scully, and Alan D'Andrea for critical reading of the manuscript.
PY - 2008/9/19
Y1 - 2008/9/19
N2 - 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.
AB - 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.
KW - DNA
UR - http://www.scopus.com/inward/record.url?scp=51549098159&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2008.08.030
DO - 10.1016/j.cell.2008.08.030
M3 - Article
C2 - 18805090
AN - SCOPUS:51549098159
SN - 0092-8674
VL - 134
SP - 969
EP - 980
JO - Cell
JF - Cell
IS - 6
ER -