Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia

Crossan, Gerry P.; Weyden, Louise van der; Rosado, Iván V.; Langevin, Frédéric; Gaillard, Patrick; McIntyre, Rebecca E; Genetics, Sanger Mouse; Gallagher, Ferdia A.; Kettunen, Mikko I.; Lewis, David Y.; Brindle, Kevin M.; Arends, Mark J.; Adams, David J.; Patel, Ketan

doi:10.1038/ng.752

Cited by 181 publications

(202 citation statements)

References 29 publications

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“…1C), indicating that SLX4 is essential for cell proliferation at least in DT40 cells. This result is marked contrast to the viability of SLX4 knockout mice (30). This difference might be attributable to the absence of the MUS81 gene in the chicken genome.…”

Section: Slx4 Is Essential For Cell Proliferationcontrasting

confidence: 51%

Involvement of SLX4 in interstrand cross-link repair is regulated by the Fanconi anemia pathway

Yamamoto

Kobayashi

Tsuda

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

180

156

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Interstrand cross-links (ICLs) block replication and transcription and thus are highly cytotoxic. In higher eukaryotes, ICLs processing involves the Fanconi anemia (FA) pathway and homologous recombination. Stalled replication forks activate the eight-subunit FA core complex, which ubiquitylates FANCD2-FANCI. Once it is posttranslationally modified, this heterodimer recruits downstream members of the ICL repairosome, including the FAN1 nuclease. However, ICL processing has been shown to also involve MUS81-EME1 and XPF-ERCC1, nucleases known to interact with SLX4, a docking protein that also can bind another nuclease, SLX1. To investigate the role of SLX4 more closely, we disrupted the SLX4 gene in avian DT40 cells. SLX4 deficiency caused cell death associated with extensive chromosomal aberrations, including a significant fraction of isochromatid-type breaks, with sister chromatids broken at the same site. SLX4 thus appears to play an essential role in cell proliferation, probably by promoting the resolution of interchromatid homologous recombination intermediates. Because ubiquitylation plays a key role in the FA pathway, and because the N-terminal region of SLX4 contains a ubiquitinbinding zinc finger (UBZ) domain, we asked whether this domain is required for ICL processing. We found that SLX4 −/− cells expressing UBZ-deficient SLX4 were selectively sensitive to ICL-inducing agents, and that the UBZ domain was required for interaction of SLX4 with ubiquitylated FANCD2 and for its recruitment to DNAdamage foci generated by ICL-inducing agents. Our findings thus suggest that ubiquitylated FANCD2 recruits SLX4 to DNA damage sites, where it mediates the resolution of recombination intermediates generated during the processing of ICLs.endonuclease | mitomycin C | cisplatin | DNA repair I nterstrand cross-links (ICLs) inhibit transcription and replication. Their considerable cytotoxicity has been ascribed primarily to their blockage of replication forks, and this phenomenon is believed to be responsible for the success of ICL-inducing agents, such as cisplatin and mitomycin-C (MMC), in cancer chemotherapy (1). ICL processing is complex, involving proteins from several distinct pathways of DNA metabolism. In higher organisms, ICL processing is orchestrated by the Fanconi anemia (FA) pathway (2, 3). Collision of replication forks with ICLs activates the ATR kinase, which in turn licenses the FANCL ubiquitin ligase subunit of the FA core complex (composed of FANCA, B, C, E, F, G, L and M proteins) to modify the FANCD2-FANCI heterodimer (2, 4-6). The monoubiquitylated FANCD2-FANCI complex is then targeted to chromatin (7,8), where it recruits downstream components of the repairosome, including the structure-specific nuclease FAN1 (9-12). However, ICL processing also requires other enzymes, such as the nucleases MUS81-EME1 and XPF-ERCC1, and how these are recruited to sites of damage in ICL repair is not known.The structure-specific endonucleases XPF-ERCC1 and MUS81-EME1, which are implicated in ICL repair and in the res...

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Section: Slx4 Is Essential For Cell Proliferationcontrasting

confidence: 51%

Involvement of SLX4 in interstrand cross-link repair is regulated by the Fanconi anemia pathway

Yamamoto

Kobayashi

Tsuda

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

180

156

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“…Another mutation in XPF's helicase‐like domain, hs G325E ( xl G314E), was reported to disrupt the interaction of XPF with the BTB domain of SLX4 (Andersen et al , 2009). This interaction is likely specific to ICL repair and not NER because SLX4‐deficient cells are not UV sensitive (Crossan et al , 2011). With the aim to further disrupt this interaction, we generated a deletion mutant lacking G314 and three surrounding residues that were predicted to form a loop (XPF ΔNSGW ).…”

Section: Resultsmentioning

confidence: 99%

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

et al. 2017

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XPF‐ERCC1 is a structure‐specific endonuclease pivotal for several DNA repair pathways and, when mutated, can cause multiple diseases. Although the disease‐specific mutations are thought to affect different DNA repair pathways, the molecular basis for this is unknown. Here we examine the function of XPF‐ERCC1 in DNA interstrand crosslink (ICL) repair. We used Xenopus egg extracts to measure both ICL and nucleotide excision repair, and we identified mutations that are specifically defective in ICL repair. One of these separation‐of‐function mutations resides in the helicase‐like domain of XPF and disrupts binding to SLX4 and recruitment to the ICL. A small deletion in the same domain supports recruitment of XPF to the ICL, but inhibited the unhooking incisions most likely by disrupting a second, transient interaction with SLX4. Finally, mutation of residues in the nuclease domain did not affect localization of XPF‐ERCC1 to the ICL but did prevent incisions on the ICL substrate. Our data support a model in which the ICL repair‐specific function of XPF‐ERCC1 is dependent on recruitment, positioning and substrate recognition.

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“…SLX4 is mutated in patients with bona fide FA (FANCP) [2,20]. In contrast to FA mouse models, the Slx4knockout (KO) mouse exhibits an FA phenotype with developmental defects and cytopenia [49]. The N-terminal segment of SLX4/FANCP harbors ubiquitin zinc finger (UBZ) domains, suggesting that SLX4 is recruited to DNA damage via interaction with ubiquitinated proteins involved in the DNA damage response (DDR).…”

Section: Fa Genes (Fanca B C D1 D2 E F G I J L N P Q) Fmentioning

confidence: 99%

Fanconi anemia: a model disease for studies on human genetics and advanced therapeutics

Bogliolo

Surrallés

2015

Current Opinion in Genetics & Development

162

167

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Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia

Cited by 181 publications

References 29 publications

Involvement of SLX4 in interstrand cross-link repair is regulated by the Fanconi anemia pathway

Involvement of SLX4 in interstrand cross-link repair is regulated by the Fanconi anemia pathway

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

Fanconi anemia: a model disease for studies on human genetics and advanced therapeutics

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