Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperaturesensitive mutant altered in DNA primase-polymerase complex stability
✍ Scribed by Lucchini, Giovanna ;Mazza, Cinzia ;Scacheri, Emanuela ;Plevani, Paolo
- Publisher
- Springer
- Year
- 1988
- Tongue
- English
- Weight
- 831 KB
- Volume
- 212
- Category
- Article
- ISSN
- 0026-8925
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✦ Synopsis
The cloned DNA polymerase I gene has been used to map the POL1 locus on the left arm of chromosome XIV, between MET4 and TOP2. Temperature-sensitive mutants in POL1 have been obtained by in vitro mutagenesis of the cloned gene and in vivo replacement of the wild-type allele with the mutated copy. Physiological and biochemical characterization of one temperature-sensitive mutant (pol1-1) shows that cells shifted to the non-permissive temperature can complete one round of cell division and DNA replication before they arrest. Analysis of DNA polymerase I in crude extracts and in partially purified preparations indicates that the pol1-1 mutation results in a conformational change and affects the stability of the DNA primase-polymerase complex.