Open AccessStudies on Deoxyribonucleic Acid Polymerases from Yeast
Author(s) -
Wintersberger Ulrike,
Wintersberger Erhard
Publication year - 1970
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1970.tb00894.x
Subject(s) - dna polymerase , polymerase , biochemistry , primer (cosmetics) , dna clamp , biology , dna polymerase ii , dna , yeast , microbiology and biotechnology , enzyme , primase , mitochondrial dna , saccharomyces cerevisiae , dna polymerase i , chemistry , rna , reverse transcriptase , gene , organic chemistry
Mitochondrial DNA polymerase from Saccharomyces cerevisiae was purified 80‐to 100‐fold with good yield by DEAE‐cellulose chromatography of the extract prepared by disruption of highly purified yeast mitochondria. The enzyme requires all four deoxyribonucleoside triphosphates, Mg ++ and DNA for activity. The optimal Mg ++ concentration is 50 mM. Different DNAs, in native as well as denatured or “activated” form, serve as template and primer. The preference for denatured over native DNA and the effective promotion of DNA synthesis by mitochondrial DNA is of special interest. Mitochondria from a cytoplasmic “petite” mutant also contain a DNA polymerase which can be solubilized and purified like the enzyme from wild type yeast. Identical properties were found for the polymerases from normal and mutant mitochondria. On the other hand, characteristic differences between the mitochondrial enzyme and the polymerases A and B obtained from mitochondria‐free cell extracts were noted in the behaviour on DEAE‐cellulose columns, the optimal Mg ++ requirements and the effectiveness of different DNA templates. DNA synthesis with all three enzymes is inhibited in a similar fashion by the intercalating dye acriflavin. The molecular weight of the mitochondrial DNA polymerase from yeast is about 150000.