Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperature-sensitive mutant altered in DNA primase-polymerase complex stability |
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Authors: | Giovanna Lucchini Cinzia Mazza Emanuela Scacheri and Paolo Plevani |
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Institution: | (1) Dipartimento di Genetica e di Biologia dei Microrganismi, Università di Milano, Via Celoria 26, I-20133 Milano, Italy |
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Abstract: | Summary 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. |
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Keywords: | Yeast DNA primase-polymerase complex Temperature sensitive mutants |
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