Coordination of Rad1–Rad10 interactions with Msh2–Msh3, Saw1 and RPA is essential for functional 3′ non-homologous tail removal
| Autor: | Robin Eichmiller, Jennifer A. Surtees, Christopher Kim, Ja Hwan Seol, Melisa Medina-Rivera, Cory Holland, Ignacio F. Gallardo, Diane Oramus, Rachel DeSanto, Jessica Smith, Eugen Minca, Sang Eun Lee, Megan Schmit, Ilya J. Finkelstein |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
DNA End-Joining Repair Saccharomyces cerevisiae Proteins Ultraviolet Rays Saccharomyces cerevisiae Genome Integrity Repair and Replication 03 medical and health sciences Endonuclease chemistry.chemical_compound Replication Protein A Protein Interaction Mapping Genetics DNA Breaks Double-Stranded biology Single-Strand Specific DNA and RNA Endonucleases Endonucleases biology.organism_classification Phenotype Cell biology DNA-Binding Proteins DNA Repair Enzymes MutS Homolog 2 Protein 030104 developmental biology chemistry MSH3 MSH2 MutS Homolog 3 Protein Mutation biology.protein DNA Nucleotide excision repair |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | Double strand DNA break repair (DSBR) comprises multiple pathways. A subset of DSBR pathways, including single strand annealing, involve intermediates with 3′ non-homologous tails that must be removed to complete repair. In Saccharomyces cerevisiae, Rad1–Rad10 is the structure-specific endonuclease that cleaves the tails in 3′ non-homologous tail removal (3′ NHTR). Rad1–Rad10 is also an essential component of the nucleotide excision repair (NER) pathway. In both cases, Rad1–Rad10 requires protein partners for recruitment to the relevant DNA intermediate. Msh2–Msh3 and Saw1 recruit Rad1–Rad10 in 3′ NHTR; Rad14 recruits Rad1–Rad10 in NER. We created two rad1 separation-of-function alleles, rad1R203A,K205A and rad1R218A; both are defective in 3′ NHTR but functional in NER. In vitro, rad1R203A,K205A was impaired at multiple steps in 3′ NHTR. The rad1R218A in vivo phenotype resembles that of msh2- or msh3-deleted cells; recruitment of rad1R218A–Rad10 to recombination intermediates is defective. Interactions among rad1R218A–Rad10 and Msh2–Msh3 and Saw1 are altered and rad1R218A–Rad10 interactions with RPA are compromised. We propose a model in which Rad1–Rad10 is recruited and positioned at the recombination intermediate through interactions, between Saw1 and DNA, Rad1–Rad10 and Msh2–Msh3, Saw1 and Msh2–Msh3 and Rad1–Rad10 and RPA. When any of these interactions is altered, 3′ NHTR is impaired. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|