A meiotic DNA polymerase from Coprinus cinereus: further purification and characterization.
A meiotic DNA polymerase that is present at a high level of activity in meiotic cells of a basidiomycete, Coprinus cinereus, was purified to near homogeneity using synthetic RNA homopolymer [poly(C)] cellulose column chromatography. This report presents the first extensive purification and characterization of any eukaryotic DNA polymerase having a role in meiosis. This enzyme is a single polypeptide with a molecular mass of 65,000. Activity in this enzyme requires magnesium ions and occurs at an optimal pH of 7.5. It is strongly inhibited by dideoxythymidine triphosphate but is relatively insensitive to aphidicolin and N-ethylmaleimide and can use poly(C)/oligo(dG)12-18 as a template-primer. Polymerase activity can be found only in cells at meiotic prophase, even though the enzyme has been identified in somatic cells in an inactive state using immunoblot analysis. Its distinctive distribution makes possible a genetic and biochemical analysis of functional role of a meiotic DNA polymerase in meiotic recombination, repair and synthesis.
Matsuda
S
S
Department of Applied Biological Science, Science University of Tokyo, Japan.
Takami
K
K
Sono
A
A
Sakaguchi
K
K
D016590:Aphidicolin / Q000494:pharmacology; D001487:Basidiomycota / Q000166:cytology / Q000201:enzymology*; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D054306:Dideoxynucleotides; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008540:Meiosis; D008970:Molecular Weight; D019384:Nucleic Acid Synthesis Inhibitors; D011119:Polynucleotides; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000494:pharmacology
D054306:Dideoxynucleotides; D019384:Nucleic Acid Synthesis Inhibitors; D011119:Polynucleotides; D013942:Thymine Nucleotides; D016590:Aphidicolin; D004259:DNA-Directed DNA Polymerase; C017216:2',3'-dideoxythymidine triphosphate; D005033:Ethylmaleimide