DNA damage triggers SAF-A and RNA biogenesis factors exclusion from chromatin coupled to R-loops removal Sébastien Britton, Emma Dernoncourt, Christine Delteil, Carine Froment, Odile Schiltz, Bernard Salles, Philippe Frit, Patrick Calsou
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Sébastien Britton, Emma Dernoncourt, Christine Delteil, Carine Froment, Odile Schiltz, et al.. DNA damage triggers SAF-A and RNA biogenesis factors exclusion from chromatin coupled to R-loops removal. Nucleic Acids Research, Oxford University Press, 2014, 42 (14), pp.9047-9062. 10.1093/nar/gku601. hal-01191434
HAL Id: hal-01191434 https://hal.archives-ouvertes.fr/hal-01191434 Submitted on 6 Oct 2015
HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Published online 16 July 2014 Nucleic Acids Research, 2014, Vol. 42, No. 14 9047–9062 doi: 10.1093/nar/gku601 DNA damage triggers SAF-A and RNA biogenesis factors exclusion from chromatin coupled to R-loops removal Sebastien´ Britton1,2,3,†, Emma Dernoncourt1,2,3,†, Christine Delteil1,2,3, Carine Froment1,2, Odile Schiltz1,2, Bernard Salles1,2, Philippe Frit1,2,3,* and Patrick Calsou1,2,3,*
1CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), BP 64182, 205 route de Narbonne, F-31077 Toulouse, Cedex 4, France, 2Universite´ de Toulouse, UPS, IPBS, F-31077 Toulouse, France and 3Equipe Labellisee´ Ligue Nationale Contre le Cancer
Received March 11, 2014; Revised June 01, 2014; Accepted June 23, 2014
ABSTRACT INTRODUCTION We previously identified the heterogeneous ribonu- Deoxyribonucleic acid (DNA) double-strand break (DSB) cleoprotein SAF-A/hnRNP U as a substrate for DNA- is the most toxic type of DNA damage. If improperly re- PK, a protein kinase involved in DNA damage re- paired, DSBs can cause cell death or mutations and gross sponse (DDR). Using laser micro-irradiation in hu- chromosomal rearrangements promoting cancer develop- man cells, we report here that SAF-A exhibits a two- ment (1–4). In mammalian cells, DSBs initiate a global DNA damage response (DDR) to overcome their toxicity phase dynamics at sites of DNA damage, with a and maintain genome stability. DDR includes lesions detec- rapid and transient recruitment followed by a pro- tion, checkpoint activation, modulation of gene expression longed exclusion. SAF-A recruitment corresponds to and DNA repair (5–9). DDR defects manifest as a variety its binding to Poly(ADP-ribose) while its exclusion of human diseases, including neurodegenerative disorders, is dependent on the activity of ATM, ATR and DNA- immunodeficiency, infertility and cancer (5). PK and reflects the dissociation from chromatin of Another component of the DDR is local transcription SAF-A associated with ongoing transcription. Hav- arrest triggered by DNA breaks (10–13). More generally, ing established that SAF-A RNA-binding domain re- an expanding aspect of the DDR is its connection with ri- capitulates SAF-A dynamics, we show that this do- bonucleic acid (RNA) metabolism. Indeed, the DNA dam- main is part of a complex comprising several mRNA age activated kinases ATM or ATR phosphorylate numer- biogenesis proteins of which at least two, FUS/TLS ous proteins involved in RNA metabolism (14,15) and links with the DDR have been established for several members of and TAFII68/TAF15, exhibit similar biphasic dynam- the heterogeneous ribonucleoprotein (hnRNP) family (16), ics at sites of damage. Using an original reporter for RNA-binding proteins (RBPs) (17–25) or pre-RNA pro- live imaging of DNA:RNA hybrids (R-loops), we show cessing factors (26,27). Moreover, RNA-processing factors a transient transcription-dependent accumulation of are major mediators of genome stability, some of them by R-loops at sites of DNA damage that is prolonged preventing interactions between the nascent RNA and tem- upon inhibition of RNA biogenesis factors exclusion. plate DNA (R-loops) (28–33) which are relevant source of We propose that a new component of the DDR is DNA breaks (33,34). an active anti-R-loop mechanism operating at dam- We and another group have identified SAF-A/hnRNP U aged transcribed sites which includes the exclusion (hereinafter referred to as SAF-A), as a substrate for DNA- of mRNA biogenesis factors such as SAF-A, FUS and PK, a key protein kinase involved in DSB repair by non- TAF15. homologous end-joining (NHEJ) (35,36). In NHEJ, DNA- PK operates together with the DSBs sensor Ku70/80 het- erodimer and the XRCC4/DNA ligase IV ligation com- plex (37). SAF-A is an abundant nuclear protein found in
*To whom correspondence should be addressed. Tel: +33 561 175 970; Fax:+33 561 175 933; Email: [email protected] Correspondence may also be addressed to Philippe Frit. Tel: +33 561 175 937; Fax:+33 561 175 933; Email: [email protected] †The authors wish it to be known that, in their opinion, the first two authors should be regarded as Joint First Authors. Present address: Bernard Salles, TOXALIM (Research Centre in Food Toxicology), UMR 1331 INRA/INP/UPS, 180 chemin de Tournefeuille, F-31027 Toulouse Cedex 3, France.