Physical interaction between SLX4 (FANCP) and XPF (FANCQ) proteins and biological consequences of interaction-defective missense mutations

Hashimoto, Keiji; Wada, Kunio; Matsumoto, Kyomu; Moriya, Masaaki

doi:10.1016/j.dnarep.2015.09.022

Cited by 15 publications

(26 citation statements)

References 25 publications

(38 reference statements)

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“…We previously showed that the xlXPF-hsERCC1 complex (referred to as XPF-ERCC1 from here on) supports ICL repair in Xenopus egg extracts (Klein Douwel et al, 2014). Expression levels of all mutant complexes were similar to wild-type levels except for the XPF L219P -ERCC1 (XE L219P ) complex that was previously reported to be unstable Hashimoto et al, 2015). To examine the function of leucine 219, we instead mutated it to an arginine and found that XE L219R was expressed at normal levels.…”

Section: Xpf Mutants Form Functional Complexes With Ercc1mentioning

confidence: 79%

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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Section: Xpf Mutants Form Functional Complexes With Ercc1mentioning

confidence: 79%

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

et al. 2017

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“…We generated a series of SLX4 deletion mutants based on SLX4’s known interaction domain or functional motif. We found that deletion of the MEI9 XPF interaction like region (MLR) of SLX4 (residues 409–555), which is considered the binding site for XPF–ERCC1 (84,85), abolished its interaction with SLX4IP (Figure 3B). We generated further small-deletion mutants in the MLR domain and found that deletion of residues 501–555 of SLX4 not only abolished its interaction with XPF but also impaired its interaction with SLX4IP (Supplemental Figure S3A).…”

Section: Resultsmentioning

confidence: 98%

“…SLX4 and XPF–ERCC1 are the major SLX4IP-interacting proteins, with or without MMC-based treatment (Figure 3A). Studies have demonstrated that SLX4 coordinates with XPF–ERCC1 and participates in the unhooking of ICLs during the repair process (31,43,67,84). The binding of SLX4IP to both SLX4 and XPF–ERCC1 inspired us to further investigate whether SLX4IP plays a role in regulating SLX4–XPF–ERCC1 interaction.…”

Section: Resultsmentioning

confidence: 99%

SLX4IP acts with SLX4 and XPF–ERCC1 to promote interstrand crosslink repair

Zhang

Chen

Yin

et al. 2019

Nucleic Acids Research

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Interstrand crosslinks (ICLs) are highly toxic DNA lesions that are repaired via a complex process requiring the coordination of several DNA repair pathways. Defects in ICL repair result in Fanconi anemia, which is characterized by bone marrow failure, developmental abnormalities, and a high incidence of malignancies. SLX4, also known as FANCP, acts as a scaffold protein and coordinates multiple endonucleases that unhook ICLs, resolve homologous recombination intermediates, and perhaps remove unhooked ICLs. In this study, we explored the role of SLX4IP, a constitutive factor in the SLX4 complex, in ICL repair. We found that SLX4IP is a novel regulatory factor; its depletion sensitized cells to treatment with ICL-inducing agents and led to accumulation of cells in the G2/M phase. We further discovered that SLX4IP binds to SLX4 and XPF–ERCC1 simultaneously and that disruption of one interaction also disrupts the other. The binding of SLX4IP to both SLX4 and XPF–ERCC1 not only is vital for maintaining the stability of SLX4IP protein, but also promotes the interaction between SLX4 and XPF–ERCC1, especially after DNA damage. Collectively, these results demonstrate a new regulatory role for SLX4IP in maintaining an efficient SLX4–XPF–ERCC1 complex in ICL repair.

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“…The second allele of XPF presented a novel splice site mutation (c.793-2A>G) which produced a premature termination of translation (p.Thr265Valfs*13), hence originating a null allele as happened with the previously reported FA XPF variants [35]. Contrary to Hashimoto group, who found the XPF-R589W mutation was abruptly affecting XPF structure and its SLX4 binding [45]. Popp and colleagues detected a residual proportion of XPF-R589W escaping from protein misfolding, able to reach the chromatin [43] as happened with the reported FA XPF variants.…”

Section: Resultsmentioning

confidence: 99%

Functional Comparison of XPF Missense Mutations Associated to Multiple DNA Repair Disorders

Marín

Ramı́rez

Aza‐Carmona

et al. 2019

Genes

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XPF endonuclease is one of the most important DNA repair proteins. Encoded by XPF/ERCC4, XPF provides the enzymatic activity of XPF-ERCC1 heterodimer, an endonuclease that incises at the 5’ side of various DNA lesions. XPF is essential for nucleotide excision repair (NER) and interstrand crosslink repair (ICLR). XPF/ERCC4 mutations are associated with several human diseases: Xeroderma Pigmentosum (XP), Segmental Progeria (XFE), Fanconi Anemia (FA), Cockayne Syndrome (CS), and XP/CS combined disease (XPCSCD). Most affected individuals are compound heterozygotes for XPF/ERCC4 mutations complicating the identification of genotype/phenotype correlations. We report a detailed overview of NER and ICLR functional studies in human XPF-KO (knock-out) isogenic cells expressing six disease-specific pathogenic XPF amino acid substitution mutations. Ultraviolet (UV) sensitivity and unscheduled DNA synthesis (UDS) assays provide the most reliable information to discern mutations associated with ICLR impairment from mutations related to NER deficiency, whereas recovery of RNA synthesis (RRS) assays results hint to a possible role of XPF in resolving R-loops. Our functional studies demonstrate that a defined cellular phenotype cannot be easily correlated to each XPF mutation. Substituted positions along XPF sequences are not predictive of cellular phenotype nor reflect a particular disease. Therefore, in addition to mutation type, allelic interactions, protein stability and intracellular distribution of mutant proteins may also contribute to alter DNA repair pathways balance leading to clinically distinct disorders.

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Physical interaction between SLX4 (FANCP) and XPF (FANCQ) proteins and biological consequences of interaction-defective missense mutations

Cited by 15 publications

References 25 publications

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

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

SLX4IP acts with SLX4 and XPF–ERCC1 to promote interstrand crosslink repair

Functional Comparison of XPF Missense Mutations Associated to Multiple DNA Repair Disorders

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