Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition

Berti, Matteo; Chaudhuri, Arnab Ray; Thangavel, Saravanabhavan; Gomathinayagam, Shivasankari; Kenig, Saša; Vujanovic, Marko; Odreman, Federico; Glatter, Timo; Graziano, Simona; Mendoza-Maldonado, Ramiro; Marino, Francesca; Lucic, Bojana; Biasin, Valentina; Gstaiger, Matthias; Aebersold, Ruedi; Sidorova, Julia M.; Monnat, Raymond J.; Lopes, Massimo; Vindigni, Alessandro

doi:10.1038/nsmb.2501

Cited by 409 publications

(517 citation statements)

References 42 publications

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“…6A). Using an optimized pulsed-field gel-electrophoresis (PFGE) protocol, which can detect Ͻ100 DSB per cell (11,12), we verified that ATM and ATR activation upon IR treatment is associated with marked accumulation of DSB (Fig. 6B) and with the expected phosphorylation of RPA32 on S4/S8 (Fig.…”

Section: Resultsmentioning

confidence: 91%

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Poly(ADP-Ribosyl) Glycohydrolase Prevents the Accumulation of Unusual Replication Structures during Unperturbed S Phase

Chaudhuri

Ahuja

Herrador

et al. 2015

Molecular and Cellular Biology

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Poly(ADP-ribosyl)ation (PAR) has been implicated in various aspects of the cellular response to DNA damage and genome stability. Although 17 human poly(ADP-ribose) polymerase (PARP) genes have been identified, a single poly(ADP-ribosyl) glycohydrolase (PARG) mediates PAR degradation. Here we investigated the role of PARG in the replication of human chromosomes. We show that PARG depletion affects cell proliferation and DNA synthesis, leading to replication-coupled H2AX phosphorylation. Furthermore, PARG depletion or inhibition per se slows down individual replication forks similarly to mild chemotherapeutic treatment. Electron microscopic analysis of replication intermediates reveals marked accumulation of reversed forks and single-stranded DNA (ssDNA) gaps in unperturbed PARG-defective cells. Intriguingly, while we found no physical evidence for chromosomal breakage, PARG-defective cells displayed both ataxia-telangiectasia-mutated (ATM) and ataxia-Rad3-related (ATR) activation, as well as chromatin recruitment of standard double-strand-break-repair factors, such as 53BP1 and RAD51. Overall, these data prove PAR degradation to be essential to promote resumption of replication at endogenous and exogenous lesions, preventing idle recruitment of repair factors to remodeled replication forks. Furthermore, they suggest that fork remodeling and restarting are surprisingly frequent in unperturbed cells and provide a molecular rationale to explore PARG inhibition in cancer chemotherapy.C ellular responses are crucial for the adaptability and survival of a cell exposed to different types of endogenous and exogenous stress. The DNA damage response (DDR) consists of one such defense mechanism in response to different types of insults to the DNA. Poly(ADP)ribosylation of proteins is one of the quickest cellular responses to DNA damage and is brought about by proteins of the poly(ADP) ribose polymerase family (PARP), mostly PARP1 (1). Upon being recruited to sites of the DNA damage, NAD ϩ is used as a substrate by PARP to synthesize negatively charged poly(ADP-ribosyl)ation (PAR) polymers onto itself and also its target proteins (1). Through this posttranslational modification, PARP targets a variety of nuclear proteins to facilitate the recruitment of DNA repair factors to sites of damage (2, 3). Accordingly, PARP-1 or PARP-2-deficient mice and mouse embryonic fibroblasts show chromosomal aberrations and various DNA repair defects (4-6).Inhibition of PARP has become a promising therapeutic approach for the treatment of certain types of cancer (7). It was shown that PARP inhibitors could selectively kill homologous recombination (HR)-deficient cancer cells (8, 9). The reason behind the sensitivity of HR-deficient cells to PARP inhibition is thought to be the accumulation of single-stranded DNA (ssDNA) breaks in the absence of PAR synthesis, leading to replication fork collapse and double-stranded breaks (DSBs), which would then require HR factors for repair (8,10). Recently, PARP activity has also been reported to play a ro...

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Section: Resultsmentioning

confidence: 91%

“…Recently, PARP activity has also been reported to play a role in the control of replication fork reversal upon topoisomerase 1 poisoning (11). Upon auto-PARylation, PARP1 interacts with the RecQ1 helicase and inhibits its specific fork restart activity, thereby transiently preventing restart of the reversed forks until repair of the damage has occurred (12).…”

mentioning

confidence: 99%

Poly(ADP-Ribosyl) Glycohydrolase Prevents the Accumulation of Unusual Replication Structures during Unperturbed S Phase

Chaudhuri

Ahuja

Herrador

et al. 2015

Molecular and Cellular Biology

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“…First, they mediate the recombination steps required to skip bulky lesions that will be processed later by specific repair mechanisms [52]. Additionally, they work on fork regression and restoration of chicken foot structures to normal forks that may be generated by lesions such as ICL [53]. In fact, the absence of these proteins increases genome instability as reflected by the expression of common fragile sites, along with increased sister chromatid exchanges and anaphase bridges [49,50,54].…”

Section: Rs As a Fuel Of Tumorogenesismentioning

confidence: 99%

“…In addition, PAR protects reversed forks from MRE11 mediated degradation by either physically impeding the access to the fork or by cooperating in the recruitment of BRCA2 and RAD51. Additionally, PARylation also blocks the access of the RECQL1 helicase and prevents the restart of the fork before it has been repaired [53]. Finally, PARylation at stalled forks also transiently recruits the scaffold protein SAFB1.…”

Section: Rs As a Fuel Of Tumorogenesismentioning

confidence: 99%

Replication stress and cancer: It takes two to tango

Lecona

Fernández-Capetillo

2014

Experimental Cell Research

119

107

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Problems arising during DNA replication require the activation of the ATR-CHK1 pathway to ensure the stabilization and repair of the forks, and to prevent the entry into mitosis with unreplicated genomes. Whereas the pathway is essential at the cellular level, limiting its activity is particularly detrimental for some cancer cells. Here we review the links between replication stress (RS) and cancer, which provide a rationale for the use of ATR and Chk1 inhibitors in chemotherapy. First, we describe how the activation of oncogene-induced RS promotes genome rearrangements and chromosome instability, both of which could potentially fuel carcinogenesis. Next, we review the various pathways that contribute to the suppression of RS, and how mutations in these components lead to increased cancer incidence and/or accelerated ageing. Finally, we summarize the evidence showing that tumours with high levels of RS are dependent on a proficient RS-response, and therefore vulnerable to ATR or Chk1 inhibitors.

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“…For example, the dynamic interplay of RECQL duplex separation and rezipping of unwound strands is likely to aid in the remodeling of stalled forks in human cancer cells exposed to the topoisomerase inhibitor camptothecin. 36 …”

Section: Molecular Analysis Of Recql Breast Cancer Associated Mutationsmentioning

confidence: 99%

RECQL: a new breast cancer susceptibility gene

Banerjee

Brosh

2015

Cell Cycle

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Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition

Cited by 409 publications

References 42 publications

Poly(ADP-Ribosyl) Glycohydrolase Prevents the Accumulation of Unusual Replication Structures during Unperturbed S Phase

Poly(ADP-Ribosyl) Glycohydrolase Prevents the Accumulation of Unusual Replication Structures during Unperturbed S Phase

Replication stress and cancer: It takes two to tango

RECQL: a new breast cancer susceptibility gene

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