Maintenance of Genome Integrity during DNA Replication

 Philippe PASERO

 IGH - UMR 9002

 141, rue de la Cardonille, 34396 Montpellier

Phone: +33 4 34 35 99 43

Email: philippe.pasero@igh.cnrs.fr

Website

DNA replication is a complex process that depends on the sequential activation of thousands of origins. Forks progressing from these origins frequently stall when they encounter obstacles such as DNA lesions or tightly-bound protein-DNA complexes. Studies in model organisms have shown that stalled forks are fragile structures that must be promptly restarted to prevent genomic instability. Fork recovery depends on various mechanisms involving specialized helicases, checkpoint kinases and DNA repair pathways. Cells mutated for these pathways show increased replication stress, unstable genomes and are prone to develop cancers. Increased replication defects are also detected in oncogene-activated cells. However, the mechanisms involved remain poorly understood. Work in our lab aims at (i) characterizing the cellular responses to replication stress and (ii) understanding the origin of spontaneous replication stress in yeast and in human cells. To this end, we use a combination of single-molecule (DNA combing) and genome-wide approaches (ChIP-chip, ChIP-seq) to monitor the dynamics of DNA replication in yeast and in human cells. These studies should bring new insights into the mechanism by which spontaneous replication stress contributes to tumorigenesis and allow the development of more efficient anticancer treatments targeting replication forks.

Keywords: DNA replication – Genomic instability – Checkpoints – Budding Yeast – Human cells

Main publications

• Tittel-Elmer M#, Lengronne A#, Davidson MB, Bacal J, François P, Hohl M, Petrini J, Pasero P* and Cobb JA* (2012) Cohesin association to replication sites depends on Rad50 and promotes fork restart. Mol Cell, 48, 98-108. (*corresponding authors)
• Poli J, Tsaponina O, Crabbe L, Keszthelyi A, Pantesco V, Chabes A, Lengronne A* and Pasero P* (2012). dNTP pools determine fork progression and origin usage under replication stress. EMBO J 31, 883-894 (*corresponding authors)
• Wurtele H, Schalck Kaiser G, Bacal J, St-Hilaire E, Lee EH, Dorn JF, Maddox P, Lisby M, Pasero P and Verreault A. (2012). Histone H3 lysine 56 acetylation and the response to DNA replication fork damage. Mol Cell Biol 32, 154-172.
• Bianco JN, Poli P, Saksouk J, Bacal J, Silva MJ, Yoshida K, Lin YL, Tourrière H, Lengronne A and Pasero P (2012) Analysis of DNA replication profiles in budding yeast and mammalian cells using DNA combing. Methods, 57, 149-157
• Lin YL and Pasero P (2012). Interference between DNA replication and transcription as a cause of genomic instability. Current Genomics, 13, 65-73
• Poli J, Tsaponina O, Crabbe L, Keszthelyi A, Pantesco V, Chabes A, Lengronne A* and Pasero P* (2011). dNTP pools determine fork progression and origin usage under replication stress. EMBO J in press. (corresponding authors)
• Crabbé L, Thomas A, Pantesco V, De Vos J, Pasero P* and Lengronne A* (2010) Genomic analysis of replication profiles identifies RFCCtf18 as a key mediator of the replication stress response. Nat Struct Mol Biol, 17, 1391-1397 (* equal contribution)
• Tuduri S, Crabbé L, Conti C, Tourrière H, Holtgreve-Grez H, Jauch A, Pantesco V, de Vos J, Theillet C, Thomas A, Pommier Y, Tazi J, Coquelle A* and Pasero P* (2009) Topoisomerase 1 suppresses replication stress and genomic instability by preventing interference between replication and transcription. Nat Cell Biol, 11, 1315-1324 (* equal contribution)
• Falbo K*, Alabert C*, Katou Y*, Wu S, Han J, Wehr T, Xiao J, He X, Zhang Z, Shi Y, Shirahige K, Pasero P and Shen X (2009) Involvement of a chromatin remodeling complex in damage tolerance during DNA replication. Nat Struct Mol Biol, 16, 1167- 1172.
• (* equal contribution)
• Alabert C, Bianco J and Pasero P (2009) Differential regulation of homologous recombination at DNA breaks and replication forks by the Mrc1 branch of the S-phase checkpoint. EMBO J, 28, 1131-1141
• Luke B, Versini G, Jaquenoud M, Waris Zaidi I, Kurz T, Pintard L, Pasero P * and Peter M * (2006) The cullin Rtt101p promotes replication fork progression through damaged DNA and natural pause sites. Curr Biol, 16, 786-792 (* corresponding authors).
• Tourrière H*, Versini G*, Cordon-Preciado V, Alabert C and Pasero P (2005) Mrc1p and Tof1p promote replication fork progression and recovery of independently of Rad53p. Mol Cell, 19, 699-706. (* equal contribution)
• Versini G, Comet I, Wu M, Hoopes L, Schwob E and Pasero P (2003) The yeast Sgs1 helicase is differentially required for genomic and ribosomal DNA replication. EMBO J. 22, 1939-1949.
• Shimada K, Pasero P and Gasser SM (2002) ORC and the intra-S phase checkpoint: a threshold model for Rad53p activation. Gene Dev. 16, 3236-3252.
• Pasero P, Bensimon A and Schwob E (2002) Single-molecule analysis reveals clustering and epigenetic regulation of replication origins at the yeast rDNA locus. Gene Dev. 16, 2479-84.