title: Combinatorial regulation of meiotic holliday junction resolution in C. elegans by HIM-6 (BLM) helicase, SLX-4, and the SLX-1, MUS-81 and XPF-1 nucleases.
abstract: Holliday junctions (HJs) are cruciform DNA structures that are created during recombination events. It is a matter of considerable importance to determine the resolvase(s) that promote resolution of these structures. We previously reported that C. elegans GEN-1 is a symmetrically cleaving HJ resolving enzyme required for recombinational repair, but we could not find an overt role in meiotic recombination. Here we identify C. elegans proteins involved in resolving meiotic HJs. We found no evidence for a redundant meiotic function of GEN-1. In contrast, we discovered two redundant HJ resolution pathways likely coordinated by the SLX-4 scaffold protein and also involving the HIM-6/BLM helicase. SLX-4 associates with the SLX-1, MUS-81 and XPF-1 nucleases and has been implicated in meiotic recombination in C. elegans. We found that C. elegans [mus-81; xpf-1], [slx-1; xpf-1], [mus-81; him-6] and [slx-1; him-6] double mutants showed a similar reduction in survival rates as slx-4. Analysis of meiotic diakinesis chromosomes revealed a distinct phenotype in these double mutants. Instead of wild-type bivalent chromosomes, pairs of "univalents" linked by chromatin bridges occur. These linkages depend on the conserved meiosis-specific transesterase SPO-11 and can be restored by ionizing radiation, suggesting that they represent unresolved meiotic HJs. This suggests the existence of two major resolvase activities, one provided by XPF-1 and HIM-6, the other by SLX-1 and MUS-81. In all double mutants crossover (CO) recombination is reduced but not abolished, indicative of further redundancy in meiotic HJ resolution. Real time imaging revealed extensive chromatin bridges during the first meiotic division that appear to be eventually resolved in meiosis II, suggesting back-up resolution activities acting at or after anaphase I. We also show that in HJ resolution mutants, the restructuring of chromosome arms distal and proximal to the CO still occurs, suggesting that CO initiation but not resolution is likely to be required for this process.
authors.affiliation: Centre for Gene Regulation and Expression, University of Dundee, Dundee, United Kingdom.
authors.forename: Ana
authors.initials: A
authors.lastname: Agostinho
authors.forename: Bettina
authors.initials: B
authors.lastname: Meier
authors.forename: Remi
authors.initials: R
authors.lastname: Sonneville
authors.forename: Marlène
authors.initials: M
authors.lastname: Jagut
authors.forename: Alexander
authors.initials: A
authors.lastname: Woglar
authors.forename: Julian
authors.initials: J
authors.lastname: Blow
authors.forename: Verena
authors.initials: V
authors.lastname: Jantsch
authors.forename: Anton
authors.initials: A
authors.lastname: Gartner
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000235:genetics*; D002843:Chromatin / Q000235:genetics; D020090:Chromosome Segregation / Q000235:genetics; D003434:Crossing Over, Genetic; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000235:genetics*; D045566:DNA, Cruciform / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D003851:Deoxyribonucleases / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics*; D006801:Humans; D008540:Meiosis / Q000235:genetics*; D049469:Meiotic Prophase I / Q000235:genetics; D051379:Mice; D009154:Mutation; D011995:Recombination, Genetic
chemical_list: D029742:Caenorhabditis elegans Proteins; D002843:Chromatin; D045566:DNA, Cruciform; D004268:DNA-Binding Proteins; C487047:HIM-6 protein, C elegans; D003851:Deoxyribonucleases; D004720:Endonucleases; C498772:MUS81 protein, human; C547401:SLX-1 protein, C elegans; C547402:XPF-1 protein, C elegans; D004265:DNA Helicases
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