Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperaturesensitive mutant altered in DNA primase-polymerase complex stability

1988 ◽  
Vol 212 (3) ◽  
pp. 459-465 ◽  
Author(s):  
Giovanna Lucchini ◽  
Cinzia Mazza ◽  
Emanuela Scacheri ◽  
Paolo Plevani
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Mapping ◽  
Dna Polymerase ◽  
Complex Stability ◽  
Dna Polymerase I ◽  
Polymerase Complex ◽  
Dna Primase ◽  
Polymerase I ◽  
I Gene

Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae

1989 ◽  
Vol 9 (10) ◽  
pp. 4447-4458
Author(s):  
T A Kunkel ◽  
R K Hamatake ◽  
J Motto-Fox ◽  
M P Fitzgerald ◽  
A Sugino
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Polymerase ◽  
Spontaneous Mutation ◽  
Catalytic Subunit ◽  
Dna Polymerase I ◽  
Spontaneous Mutation Rate ◽  
Single Base ◽  
Dna Primase ◽  
Polymerase I ◽  
Primase Complex

We have determined the fidelity of DNA synthesis by DNA polymerase I (yPol I) from Saccharomyces cerevisiae. To determine whether subunits other than the polymerase catalytic subunit influence fidelity, we measured the accuracy of yPol I purified by conventional procedures, which yields DNA polymerase with a partially proteolyzed catalytic subunit and no associated primase activity, and that of yPol I purified by immunoaffinity chromatography, which yields polymerase having a single high-molecular-weight species of the catalytic subunit, as well as three additional polypeptides and DNA primase activity. In assays that score polymerase errors within the lacZ alpha-complementation gene in M13mp2 DNA, yPol I and the yPol I-primase complex produced single-base substitutions, single-base frameshifts, and larger deletions. For specific errors and template positions, the two forms of polymerase exhibited differences in fidelity that could be as large as 10-fold. Nevertheless, results for the overall error frequency and the spectrum of errors suggest that the yPol I-DNA primase complex is not highly accurate and that, just as for the polymerase alone, its fidelity is not sufficient to account for a low spontaneous mutation rate in vivo. The specificity data also suggest models to explain -1 base frameshifts in nonrepeated sequences and certain complex deletions by a direct repeat mechanism involving aberrant loop-back synthesis.

scholarly journals Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae.

1989 ◽  
Vol 9 (10) ◽  
pp. 4447-4458 ◽  
Author(s):  
T A Kunkel ◽  
R K Hamatake ◽  
J Motto-Fox ◽  
M P Fitzgerald ◽  
A Sugino
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Polymerase ◽  
Spontaneous Mutation ◽  
Catalytic Subunit ◽  
Dna Polymerase I ◽  
Spontaneous Mutation Rate ◽  
Single Base ◽  
Dna Primase ◽  
Polymerase I ◽  
Primase Complex

We have determined the fidelity of DNA synthesis by DNA polymerase I (yPol I) from Saccharomyces cerevisiae. To determine whether subunits other than the polymerase catalytic subunit influence fidelity, we measured the accuracy of yPol I purified by conventional procedures, which yields DNA polymerase with a partially proteolyzed catalytic subunit and no associated primase activity, and that of yPol I purified by immunoaffinity chromatography, which yields polymerase having a single high-molecular-weight species of the catalytic subunit, as well as three additional polypeptides and DNA primase activity. In assays that score polymerase errors within the lacZ alpha-complementation gene in M13mp2 DNA, yPol I and the yPol I-primase complex produced single-base substitutions, single-base frameshifts, and larger deletions. For specific errors and template positions, the two forms of polymerase exhibited differences in fidelity that could be as large as 10-fold. Nevertheless, results for the overall error frequency and the spectrum of errors suggest that the yPol I-DNA primase complex is not highly accurate and that, just as for the polymerase alone, its fidelity is not sufficient to account for a low spontaneous mutation rate in vivo. The specificity data also suggest models to explain -1 base frameshifts in nonrepeated sequences and certain complex deletions by a direct repeat mechanism involving aberrant loop-back synthesis.

Mechanism of initiation of in vitro DNA synthesis by the immunopurified complex between yeast DNA polymerase I and DNA primase

1986 ◽  
Vol 161 (2) ◽  
pp. 435-440 ◽  
Author(s):  
Gianfranco BADARACCO ◽  
Paola VALSASNINI ◽  
Marco FOIANI ◽  
Roberta BENFANTE ◽  
Giovanna LUCCHINI ◽  
...  
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Dna Polymerase I ◽  
Dna Primase ◽  
Polymerase I

scholarly journals DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (12) ◽  
pp. 2811-2817 ◽  
Author(s):  
M A Resnick ◽  
A Sugino ◽  
J Nitiss ◽  
T Chow
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
High Efficiency ◽  
Intragenic Recombination ◽  
Dna Polymerase I ◽  
Crude Extracts ◽  
Total Dna ◽  
Polymerase I ◽  
Kinetics Of

We utilized strains of Saccharomyces cerevisiae that exhibit high efficiency of synchrony of meiosis to examine several aspects of meiosis including sporulation, recombination, DNA synthesis, DNA polymerase I and II, and Mg2+-dependent alkaline DNases. The kinetics of commitment to intragenic recombination and sporulation are similar. The synthesis of DNA, as measured directly with diphenylamine, appears to precede the commitment to recombination. Both DNA polymerase I and II activities and total DNA-synthesizing activity in crude extracts increase two- to threefold before the beginning of meiotic DNA synthesis. Increases of 10- to 20-fold over mitotic levels are found for Mg2+-dependent alkaline DNase activity in crude extracts before and during the commitment to meiotic intragenic recombination. Of particular interest is the comparable increase in a nuclease under the control of the RAD52 gene; this enzyme has been identified by the use of antibody raised against a similar enzyme from Neurospora crassa. Since the RAD52 gene is essential for meiotic recombination, the nuclease is implicated in the high levels of recombination observed during meiosis. The effects observed in this report are meiosis specific since they are not observed in an alpha alpha strain.

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