Structural and Mechanistic Analysis of the Slx1-Slx4 Endonuclease

Gaur, Vineet; Wyatt, Haley D.M.; Komorowska, Weronika; Szczepanowski, Roman H.; Sanctis, Daniele de; Gorecka, Karolina M.; West, Stephen C.; Nowotny, Marcin

doi:10.1016/j.celrep.2015.02.019

Cited by 30 publications

(74 citation statements)

References 45 publications

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“…As a control, we generated an SMX mutant containing catalytic mutations in SLX1 (R41A and E82A), MUS81 (D307A), and XPF (D705A). These mutations impaired the activities of SMX (Figure S2E), as observed previously with the individual heterodimers (Enzlin and Schärer, 2002, Gaur et al., 2015, Wyatt et al., 2013). …”

Section: Resultssupporting

confidence: 87%

The SMX DNA Repair Tri-nuclease

et al. 2017

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SummaryThe efficient removal of replication and recombination intermediates is essential for the maintenance of genome stability. Resolution of these potentially toxic structures requires the MUS81-EME1 endonuclease, which is activated at prometaphase by formation of the SMX tri-nuclease containing three DNA repair structure-selective endonucleases: SLX1-SLX4, MUS81-EME1, and XPF-ERCC1. Here we show that SMX tri-nuclease is more active than the three individual nucleases, efficiently cleaving replication forks and recombination intermediates. Within SMX, SLX4 co-ordinates the SLX1 and MUS81-EME1 nucleases for Holliday junction resolution, in a reaction stimulated by XPF-ERCC1. SMX formation activates MUS81-EME1 for replication fork and flap structure cleavage by relaxing substrate specificity. Activation involves MUS81’s conserved N-terminal HhH domain, which mediates incision site selection and SLX4 binding. Cell cycle-dependent formation and activation of this tri-nuclease complex provides a unique mechanism by which cells ensure chromosome segregation and preserve genome integrity.

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Section: Resultssupporting

confidence: 87%

The SMX DNA Repair Tri-nuclease

et al. 2017

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“…The interaction of Slx4 with Slx1 is mediated by Slx4’s HtH domain (Fig. 2) and a recent structural study in Candida glabrata showed that the interaction is responsible for dissociating an inactive Slx1 homodimer to promote its endonuclease activity (Gaur et al 2015). The importance of Slx1-Slx4, especially in the absence of Sgs1, is likely associated with the fact that replication forks stall with high frequency at the rDNA locus due to the presence of a polar replication fork barrier (RFB) (Brewer and Fangman 1988; Linskens and Huberman 1988).…”

Section: Slx4 As a Scaffold For Coordination Of Structure-specific Enmentioning

confidence: 98%

Slx4 scaffolding in homologous recombination and checkpoint control: lessons from yeast

et al. 2016

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Homologous recombination-mediated DNA repair is essential for maintaining genome integrity. It is a multi-step process that involves resection of DNA ends, strand invasion, DNA synthesis and/or DNA end ligation and, finally, the processing of recombination intermediates such as Holliday junctions or other joint molecules. Over the last 15 years, it has been established that the Slx4 protein plays key roles in the processing of recombination intermediates, functioning as a scaffold to coordinate the action of structure-specific endonucleases. Recent work in budding yeast has uncovered unexpected roles for Slx4 in the initial step of DNA-end resection and in the modulation of DNA damage checkpoint signaling. Here we review these latest findings and discuss the emerging role of yeast Slx4 as an important coordinator of DNA damage signaling responses and a regulator of multiple steps in homologous recombination-mediated repair.

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“…Each species can be derived from the four-way junction 3 (see Figure S4), and include a common strand (the x strand) that was radioactively [5'- 32 The GIG-YIG motif of human ANKLE1 is required for nuclease cleavage close to DNA branchpoints ANKLE1 has a highly conserved GIY-YIG domain at its C-terminal end ( Figure 4A). GIY-YIG domains function as the active centers of a number of nucleases (33), including R-Eco29kl (34), I-TevI (35), UvrC (36) and the junction-resolving enzyme SLX1 (37). For ANKLE1 the sequence is FTY 453 (31 amino acids) Y 486 VG.…”

Section: Ankle1 Cleaves Double-stranded Dna Close To Branchpointsmentioning

confidence: 99%

Human ANKLE1 is a nuclease specific for branched DNA

Song

Freeman

Knebel

et al. 2020

Preprint

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ABSTRACTAll physical connections between sister chromatids must be broken before cells can divide, and eukaryotic cells have evolved multiple ways in which to process branchpoints connecting DNA molecules separated both spatially and temporally. A single DNA link between chromatids has the potential to disrupt cell cycle progression and genome integrity, so it is highly likely that cells require a nuclease that can process remaining unresolved and hemi-resolved DNA junctions and other branched species at the very late stages of mitosis. We argue that ANKLE1 probably serves this function in human cells (LEM-3 in C. elegans). LEM-3 has previously been shown to be located at the cell mid-body, and we show here that human ANKLE1 is a nuclease that cleaves a range of branched DNA species. It thus has the substrate selectivity consistent with an enzyme required to process a variety of unresolved and hemi-resolved branchpoints in DNA. Our results imply that ANKLE1 acts as a catch-all enzyme of last resort that allows faithful chromosome segregation and cell division to occur.

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Structural and Mechanistic Analysis of the Slx1-Slx4 Endonuclease

Cited by 30 publications

References 45 publications

The SMX DNA Repair Tri-nuclease

The SMX DNA Repair Tri-nuclease

Slx4 scaffolding in homologous recombination and checkpoint control: lessons from yeast

Human ANKLE1 is a nuclease specific for branched DNA

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