A method to select for mutator DNA polymerase δs inSaccharomyces cerevisiae
| Autor: | Kelly Murphy, Elizabeth Fidalgo da Silva, Amy Schultz, Linda J. Reha-Krantz, Hariyanto Darmawan |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Exonuclease
Saccharomyces cerevisiae Proteins DNA Repair Base Pair Mismatch DNA polymerase DNA polymerase II DNA Mutational Analysis Molecular Sequence Data DNA-Directed DNA Polymerase Viral Proteins Genetics Point Mutation Amino Acid Sequence Selection Genetic Molecular Biology DNA Polymerase III DNA clamp biology DNA replication General Medicine biology.protein Proofreading Primase Sequence Alignment DNA polymerase mu Cadmium Biotechnology |
| Zdroj: | Genome. 49:403-410 |
| ISSN: | 1480-3321 0831-2796 |
| DOI: | 10.1139/g05-106 |
| Popis: | Proofreading DNA polymerases share common short peptide motifs that bind Mg2+in the exonuclease active center; however, hydrolysis rates are not the same for all of the enzymes, which indicates that there are functional and likely structural differences outside of the conserved residues. Since structural information is available for only a few proofreading DNA polymerases, we developed a genetic selection method to identify mutant alleles of the POL3 gene in Saccharomyces cerevisiae, which encode DNA polymerase δ mutants that replicate DNA with reduced fidelity. The selection procedure is based on genetic methods used to identify "mutator" DNA polymerases in bacteriophage T4. New yeast DNA polymerase δ mutants were identified, but some mutants expected from studies of the phage T4 DNA polymerase were not detected. This would indicate that there may be important differences in the proofreading pathways catalyzed by the two DNA polymerases.Key words: DNA polymerase proofreading, genetic selection for mutator mutants, fidelity of DNA replication, yeast. |
| Databáze: | OpenAIRE |
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