FANCD2-associated Nuclease 1, but Not Exonuclease 1 or Flap Endonuclease 1, Is Able to Unhook DNA Interstrand Cross-links in Vitro

Pizzolato, Julia; Mukherjee, Shivam; Schärer, Orlando D.; Jiricny, Josef

doi:10.1074/jbc.m115.663666

Cited by 38 publications

(36 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, in vitro biochemical analyses of SLX4/FANCP-and XPF/ FANCQ-dependent processing of the cross-links and the ICL unhooking studies from Xenopus egg extracts strongly support the major involvement of the 3 ′ flap endonuclease activity of XPF/FANCQ in initiating and completing ICL incisions (Douwel et al 2014;Hodskinson et al 2014). However, it is evident from the published biochemical work and our unpublished data that FAN1, through its endonuclease and exonuclease activities, is able to incise and completely unhook different types of cross-link substrates in vitro (Zhao et al 2014;Pizzolato et al 2015;A Wang, L Timashev, and A Smogorzewska, unpubl.). The discrepancy may come from the fact that FAN1 may be involved at stalled forks that do not look like the divergent replication forks formed on plasmids in the Xenopus egg extracts or that FAN1 predominantly works in G1 and not in S phase.…”

Section: Fan1 Vs Other Nucleasesmentioning

confidence: 59%

“…In some cases, the processing of crosslink substrates by FAN1 occurred at both the 5 ′ and 3 ′ sides of the cross-link, leading to complete unhooking of the DNA cross-link (Wang et al 2014;Pizzolato et al 2015). To date, SNM1A, a homolog of yeast Pso2, is the only other known ICL-processing nuclease that is capable of digesting past the ICL, leading to the insight that FAN1 may be functionally redundant with SNM1A in DNA ICL repair.…”

Section: Fan1 Deficiency Can Be Partially Compensated For By Snm1amentioning

confidence: 99%

“…Regardless of the genetic interaction with the FA pathway, recent in vitro studies have highlighted a possible mode of FAN1 activity on the cross-linked DNA substrates (Wang et al 2014;Zhao et al 2014;Pizzolato et al 2015). These studies showed that FAN1 nuclease activity is able to process the ICL-stalled replication forklike structures by cleaving 5 ′ flaps and also by processing nicks made by other nucleases to digest 5 ′ to 3 ′ across the cross-link, de facto unhooking it.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Fan1 deficiency results in DNA interstrand cross-link repair defects, enhanced tissue karyomegaly, and organ dysfunction

et al. 2016

View full text Add to dashboard Cite

Deficiency of FANCD2/FANCI-associated nuclease 1 (FAN1) in humans leads to karyomegalic interstitial nephritis (KIN), a rare hereditary kidney disease characterized by chronic renal fibrosis, tubular degeneration, and characteristic polyploid nuclei in multiple tissues. The mechanism of how FAN1 protects cells is largely unknown but is thought to involve FAN1's function in DNA interstrand cross-link (ICL) repair. Here, we describe a Fan1-deficient mouse and show that FAN1 is required for cellular and organismal resistance to ICLs. We show that the ubiquitinbinding zinc finger (UBZ) domain of FAN1, which is needed for interaction with FANCD2, is not required for the initial rapid recruitment of FAN1 to ICLs or for its role in DNA ICL resistance. Epistasis analyses reveal that FAN1 has cross-link repair activities that are independent of the Fanconi anemia proteins and that this activity is redundant with the 5 ′ -3 ′ exonuclease SNM1A. Karyomegaly becomes prominent in kidneys and livers of Fan1-deficient mice with age, and mice develop liver dysfunction. Treatment of Fan1-deficient mice with ICL-inducing agents results in pronounced thymic and bone marrow hypocellularity and the disappearance of c-kit + cells. Our results provide insight into the mechanism of FAN1 in ICL repair and demonstrate that the Fan1 mouse model effectively recapitulates the pathological features of human FAN1 deficiency.

show abstract

Section: Fan1 Vs Other Nucleasesmentioning

confidence: 59%

Section: Fan1 Deficiency Can Be Partially Compensated For By Snm1amentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Fan1 deficiency results in DNA interstrand cross-link repair defects, enhanced tissue karyomegaly, and organ dysfunction

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Recently, the FAN1 nuclease has also been demonstrated to act in an exo‐like fashion and is able to degrade past and release DNA substrates containing ICLs (Wang et al , 2014; Zhao et al , 2014; Pizzolato et al , 2015). Moreover, studies in fission yeast and more recently in mouse cells imply that FAN1 and SNM1A may play a redundant role in ICL repair, or contribute to different ICL repair sub‐pathways (Fontebasso et al , 2013; Thongthip et al , 2016).…”

Section: Discussionmentioning

confidence: 99%

RPA activates the XPF ‐ ERCC 1 endonuclease to initiate processing of DNA interstrand crosslinks

et al. 2017

View full text Add to dashboard Cite

During replication‐coupled DNA interstrand crosslink (ICL) repair, the XPF‐ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF‐ERCC1 incises simple ICL‐containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single‐stranded DNA (ssDNA)‐binding replication protein A (RPA) selectively restores XPF‐ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF‐ERCC1‐RPA‐induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF‐ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

show abstract

“…In vitro studies showed that human FAN1 can degrade ICL-containing oligonucleotides past the lesion in replication-like intermediates, thereby unhooking the ICLs (32)(33)(34).…”

Section: Dna Synthesis (Tls) and The Second Strand Is Repaired By Hommentioning

confidence: 99%

Structural mechanism of DNA interstrand cross-link unhooking by the bacterial FAN1 nuclease

Jin

Roy

Lee

et al. 2018

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The mechanism of ICL unhooking by bacterial FAN1 4 a "head to tail" dimer in the presence of DNA (34).The authors suggest that FAN1 dimer scans, latches, and unwinds DNA to pull the single-stranded DNA to the active site of a FAN1. Mutations of the residues at the dimeric interface abolished the nuclease activity of the HsFAN1, raising the possibility that the dimerization is important for its activity.Bacterial FAN1 homologs share a high similarity with HsFAN1 (Fig. 1A, 23,35). Second, PaFAN1 does not have a specific basic pocket that is critical for the proposed "3-nt exonuclease" mechanism (Fig. 1B, C, 33). Third, PaFAN1 lacks the basic motif in the SAP domain required for dimerization in one of the structures of the human nuclease (Fig. 1A, 34). These features raise the question if PaFAN1 can unhook ICLs, and if so, by which mechanism.To address these issues, we examined the ICL Based on these findings, we provide insights into how FAN1 might have evolved to have two basic regions to guide its nuclease activity and maintain genomic stability. RESULTS Substrate specificity of PaFAN1In previous study, we have used a DNA substrate with a four nucleotide 5' flap (23). (Fig. S1E-F). PaFAN1 efficiently incises ICL lesionTo examine whether the bacterial FAN1homolog shares the ICL resolving activity of mammalian FAN1, we examined the in vitro ICL unhooking activity of PaFAN1 using various DNA ICL substrates (Fig. 1D). We first analyzed ifPaFAN1 can unhook the ICL near the ss/ds junction.We used DNA substrates containing a nitrogen-like ICL, which is free of duplex distortion (15,36,37), The mechanism of ICL unhooking by bacterial FAN1 7 how PaFAN1 can catalyze multiple cleavage events.

show abstract

FANCD2-associated Nuclease 1, but Not Exonuclease 1 or Flap Endonuclease 1, Is Able to Unhook DNA Interstrand Cross-links in Vitro

Cited by 38 publications

References 37 publications

Fan1 deficiency results in DNA interstrand cross-link repair defects, enhanced tissue karyomegaly, and organ dysfunction

Fan1 deficiency results in DNA interstrand cross-link repair defects, enhanced tissue karyomegaly, and organ dysfunction

RPA activates the XPF ‐ ERCC 1 endonuclease to initiate processing of DNA interstrand crosslinks

Structural mechanism of DNA interstrand cross-link unhooking by the bacterial FAN1 nuclease

Contact Info

Product

Resources

About