TY - JOUR
T1 - Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition
AU - Berti, Matteo
AU - Chaudhuri, Arnab Ray
AU - Thangavel, Saravanabhavan
AU - Gomathinayagam, Shivasankari
AU - Kenig, Sasa
AU - Vujanovic, Marko
AU - Odreman, Federico
AU - Glatter, Timo
AU - Graziano, Simona
AU - Mendoza-Maldonado, Ramiro
AU - Marino, Francesca
AU - Lucic, Bojana
AU - Biasin, Valentina
AU - Gstaiger, Matthias
AU - Aebersold, Ruedi
AU - Sidorova, Julia M.
AU - Monnat, Raymond J.
AU - Lopes, Massimo
AU - Vindigni, Alessandro
N1 - Funding Information:
We are grateful to A. Mazin for sharing information regarding the substrate preparation. We thank P. Janscak (University of Zurich) and D. Orren (University of Kentucky College of Medicine) for providing aliquots of purified WRN and WRN-E84A proteins, respectively. We thank G. de Murcia (École Supérieure de Biotechnologie de Strasbourg) for providing the constructs for the production of the PARP1 fragment. We thank Y. Ayala for critical discussions and G. Triolo for help in recombinant protein production. We thank the Nano Research Facility of the School of Engineering and Applied Science at Washington University in St. Louis, which is part of the National Nanotechnology Infrastructure Network supported by the US National Science Foundation under grant no. ECS-0335765, for microfabrication and the use of clean-room facility. We also thank the Center for Microscopy and Image Analysis of the University of Zurich for technical assistance with EM. This work was supported by startup funding from the Doisy Department of Biochemistry and Molecular Biology at the Saint Louis University School of Medicine and the Saint Louis University Cancer Center and grants from the President’s Research Fund of Saint Louis University and the Associazione Italiana per la Ricerca sul Cancro (AIRC10510) to A.V.; US National Institutes of Health grant CA77852 to R.J.M. Jr.; Swiss National Science Foundation grants PP0033-114922 and PP00P3-135292 to M.L.; and a contribution from Fonds zur Förderung des Akademischen Nachwuchses (FAN) of the Zürcher Universitätsverein (ZUNIV) to M.L. and A.R.C.
PY - 2013/3
Y1 - 2013/3
N2 - Topoisomerase I (TOP1) inhibitors are an important class of anticancer drugs. The cytotoxicity of TOP1 inhibitors can be modulated by replication fork reversal through a process that requires poly(ADP-ribose) polymerase (PARP) activity. Whether regressed forks can efficiently restart and what factors are required to restart fork progression after fork reversal are still unknown. We have combined biochemical and EM approaches with single-molecule DNA fiber analysis to identify a key role for human RECQ1 helicase in replication fork restart after TOP1 inhibition that is not shared by other human RecQ proteins. We show that the poly(ADP-ribosyl)ation activity of PARP1 stabilizes forks in the regressed state by limiting their restart by RECQ1. These studies provide new mechanistic insights into the roles of RECQ1 and PARP in DNA replication and offer molecular perspectives to potentiate chemotherapeutic regimens based on TOP1 inhibition.
AB - Topoisomerase I (TOP1) inhibitors are an important class of anticancer drugs. The cytotoxicity of TOP1 inhibitors can be modulated by replication fork reversal through a process that requires poly(ADP-ribose) polymerase (PARP) activity. Whether regressed forks can efficiently restart and what factors are required to restart fork progression after fork reversal are still unknown. We have combined biochemical and EM approaches with single-molecule DNA fiber analysis to identify a key role for human RECQ1 helicase in replication fork restart after TOP1 inhibition that is not shared by other human RecQ proteins. We show that the poly(ADP-ribosyl)ation activity of PARP1 stabilizes forks in the regressed state by limiting their restart by RECQ1. These studies provide new mechanistic insights into the roles of RECQ1 and PARP in DNA replication and offer molecular perspectives to potentiate chemotherapeutic regimens based on TOP1 inhibition.
UR - http://www.scopus.com/inward/record.url?scp=84875220657&partnerID=8YFLogxK
U2 - 10.1038/nsmb.2501
DO - 10.1038/nsmb.2501
M3 - Article
C2 - 23396353
AN - SCOPUS:84875220657
SN - 1545-9993
VL - 20
SP - 347
EP - 354
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 3
ER -