Alleles of the Yeast PMS1 Mismatch-Repair Gene That Differentially Affect Recombination- and Replication-Related Processes

• Published in: Genetics (Austin) Vol. 162; no. 3; pp. 1131 - 1145
• Main Authors: Welz-Voegele, Caroline, Stone, Jana E, Tran, Phuoc T, Kearney, Hutton M, Liskay, R. Michael, Petes, Thomas D, Jinks-Robertson, Sue
• Format: Journal Article
• Language: English
• Published: United States Genetics Soc America 01-11-2002
• Subjects: Adaptor Proteins, Signal Transducing; Adenosine Triphosphate - metabolism; Base Pair Mismatch; Carrier Proteins - genetics; Carrier Proteins - metabolism; DNA Repair; DNA Replication; Fungal Proteins - genetics; Fungal Proteins - metabolism; Mutation; MutL Protein Homolog 1; MutL Proteins; Protein Structure, Tertiary; Recombination, Genetic; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; Sequence Analysis, DNA; Two-Hybrid System Techniques
• ISSN: 0016-6731; 1943-2631; 1943-2631
• DOI: 10.1093/genetics/162.3.1131

Mismatch-repair (MMR) systems promote eukaryotic genome stability by removing errors introduced during DNA replication and by inhibiting recombination between nonidentical sequences (spellchecker and antirecombination activities, respectively). Following a common mismatch-recognition step effected by MutS-homologous Msh proteins, homologs of the bacterial MutL ATPase (predominantly the Mlh1p-Pms1p heterodimer in yeast) couple mismatch recognition to the appropriate downstream processing steps. To examine whether the processing steps in the spellchecker and antirecombination pathways might differ, we mutagenized the yeast PMS1 gene and screened for mitotic separation-of-function alleles. Two alleles affecting only the antirecombination function of Pms1p were identified, one of which changed an amino acid within the highly conserved ATPase domain. To more specifically address the role of ATP binding/hydrolysis in MMR-related processes, we examined mutations known to compromise the ATPase activity of Pms1p or Mlh1p with respect to the mitotic spellchecker and antirecombination activities and with respect to the repair of mismatches present in meiotic recombination intermediates. The results of these analyses confirm a differential requirement for the Pms1p ATPase activity in replication vs. recombination processes, while demonstrating that the Mlh1p ATPase activity is important for all examined MMR-related functions.