Sensitivity to phosphonoacetic acid: a new phenotype to probe DNA polymerase delta in Saccharomyces cerevisiae.

A mutant allele (pol3-L612M) of the DNA polymerase delta gene in Saccharomyces cerevisiae that confers sensitivity to the antiviral drug phosphonoacetic acid (PAA) was constructed. We report that PAA-sensitivity tagging DNA polymerases is a useful method for selectively and reversibly inhibiting one type of DNA polymerase. Our initial studies reveal ...
that replication by the L612M-DNA pol delta requires Rad27 flap endonuclease activity since the pol3-L612M strain is not viable in the absence of RAD27 function. The L612M-DNA pol delta also strongly depends on mismatch repair (MMR). Reduced viability is observed in the absence of any of the core MMR proteins-Msh2, Mlh1, or Pms1-and severe sensitivity to PAA is observed in the absence of the core proteins Msh6 or Exo1, but not Msh3. We propose that pol3-L612M cells need the Rad27 flap endonuclease and MMR complexes composed of Msh2/Msh6, Mlh1/Pms1, and Exo1 for correct processing of Okazaki fragments.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Alleles, Amino Acid Motifs, Base Pair Mismatch, Carrier Proteins, Cell Survival, Culture Media, DNA Mutational Analysis, DNA Polymerase III, DNA Repair, DNA-Binding Proteins, DNA-Directed DNA Polymerase, Exodeoxyribonucleases, Flap Endonucleases, Fungal Proteins, Genetic Techniques, MutS Homolog 2 Protein, Mutation, Phenotype, Phosphonoacetic Acid, Polymerase Chain Reaction, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Time Factors
Date: Jun. 01, 2005
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