title: The conserved Fanconi anemia nuclease Fan1 and the SUMO E3 ligase Pli1 act in two novel Pso2-independent pathways of DNA interstrand crosslink repair in yeast.
abstract: DNA interstrand cross-links (ICLs) represent a physical barrier to the progression of cellular machinery involved in DNA metabolism. Thus, this type of adduct represents a serious threat to genomic stability and as such, several DNA repair pathways have evolved in both higher and lower eukaryotes to identify this type of damage and restore the integrity of the genetic material. Human cells possess a specialized ICL-repair system, the Fanconi anemia (FA) pathway. Conversely yeasts rely on the concerted action of several DNA repair systems. Recent work in higher eukaryotes identified and characterized a novel conserved FA component, FAN1 (Fanconi anemia-associated nuclease 1, or FANCD2/FANCI-associated nuclease 1). In this study, we characterize Fan1 in the yeast Schizosaccharomyces pombe. Using standard genetics, we demonstrate that Fan1 is a key component of a previously unidentified ICL-resolution pathway. Using high-throughput synthetic genetic arrays, we also demonstrate the existence of a third pathway of ICL repair, dependent on the SUMO E3 ligase Pli1. Finally, using sequence-threaded homology models, we predict and validate key residues essential for Fan1 activity in ICL repair.
authors.affiliation: Genome Damage and Stability Centre, University of Sussex, Brighton, East Sussex BN1 9RQ, UK; Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
authors.forename: Y
authors.initials: Y
authors.lastname: Fontebasso
authors.forename: T J
authors.initials: TJ
authors.lastname: Etheridge
authors.forename: A W
authors.initials: AW
authors.lastname: Oliver
authors.forename: J M
authors.initials: JM
authors.lastname: Murray
authors.forename: A M
authors.initials: AM
authors.lastname: Carr
mesh_terms: D004249:DNA Damage; D004260:DNA Repair; D004706:Endodeoxyribonucleases / Q000378:metabolism; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D057166:High-Throughput Screening Assays; D008025:Ligases; D008958:Models, Molecular; D059645:Mutation Rate; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017385:Sequence Homology; D015398:Signal Transduction / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D029702:Schizosaccharomyces pombe Proteins; D044767:Ubiquitin-Protein Ligases; D004706:Endodeoxyribonucleases; D004950:Esterases; C000589459:Fanconi anemia-associated nuclease 1, S pombe; C445561:Rad13 protein, S pombe; D008025:Ligases; C587432:Pli1 protein, S pombe
keywords: Cisplatin; Epistasis; Genetic screen; ICL; Schizosaccharomyces pombe; Synthetic array