title: Genetic and physical interactions between the yeast ELG1 gene and orthologs of the Fanconi anemia pathway.
abstract: Fanconi anemia (FA) is a human syndrome characterized by genomic instability and increased incidence of cancer. FA is a genetically heterogeneous disease caused by mutations in at least 15 different genes; several of these genes are conserved in the yeast Saccharomyces cerevisiae. Elg1 is also a conserved protein that forms an RFC-like complex, which interacts with SUMOylated PCNA. The mammalian Elg1 protein has been recently found to interact with the FA complex. Here we analyze the genetic interactions between elg1Δ and mutants of the yeast FA-like pathway. We show that Elg1 physically contacts the Mhf1/Mhf2 histone-like complex and genetically interacts with MPH1 (ortholog of the FANCM helicase) and CHL1 (ortholog of the FANCJ helicase) genes. We analyze the sensitivity of double, triple, quadruple and quintuple mutants to methylmethane sulfonate (MMS) and to hydroxyurea (HU). Our results show that genetic interactions depend on the type of DNA damaging agent used and show a hierarchy: Chl1 and Elg1 play major roles in the survival to these genotoxins and exhibit synthetic fitness reduction. Mph1 plays a lesser role, and the effect of the Mhf1/2 complex is seen only in the absence of Elg1 on HU-containing medium. Finally, we dissect the relationship between yeast FA-like mutants and the replication clamp, PCNA. Our results point to an intricate network of interactions rather than a single, linear repair pathway.
authors.affiliation: Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Shivani
authors.initials: S
authors.lastname: Singh
authors.forename: Keren
authors.initials: K
authors.lastname: Shemesh
authors.forename: Batia
authors.initials: B
authors.lastname: Liefshitz
authors.forename: Martin
authors.initials: M
authors.lastname: Kupiec
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects; D005199:Fanconi Anemia / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation; D020798:Two-Hybrid System Techniques
chemical_list: C066751:CHL1 protein, S cerevisiae; D002352:Carrier Proteins; D002868:Chromosomal Proteins, Non-Histone; C477609:Elg1 protein, S cerevisiae; C572609:MHF2 protein, S cerevisiae; D018809:Proliferating Cell Nuclear Antigen; D029701:Saccharomyces cerevisiae Proteins; D008741:Methyl Methanesulfonate; C491240:MPH1 protein, S cerevisiae; D053487:DEAD-box RNA Helicases; D006918:Hydroxyurea
keywords: DNA damage; Elg1; Fanconi anemia; PCNA; genome stability