DNA polymerase epsilon from Drosophila melanogaster

Biochem Biophys Res Commun. 1997 Jan 13;230(2):297-301. doi: 10.1006/bbrc.1996.5945.

Abstract

We identified a DNA polymerase species in Drosophila melanogaster embryos, and purified it. This polymerase shared some common properties with DNA polymerase epsilon from mammals and yeast as follows; it has a preference for poly(dA)/oligo(dT) as a template/primer, it is highly processive in DNA synthesis, it co-fractionates with 3'-5' exonuclease activity, it is sensitive to aphidicolin and is resistance to ddTTP. The polymerase activity was inhibited in the immuno-precipitation assay with anti-pol-epsilon antibodies, which were produced against a polypeptide coded on the cDNA of a putative Drosophila pol-epsilon we isolated previously. Using these antibodies, Western blot analysis revealed that this polymerase is a 250kDa polypeptide, which is the same size as observed in mammals and yeast. These results indicate that Drosophila produces the epsilon-class of DNA polymerase, and like mammals or yeast, possesses the 5 typical classes of DNA polymerases (alpha to epsilon) in its embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aphidicolin / pharmacology
  • Blotting, Western
  • Chromatography
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Cytosol / enzymology
  • DNA Polymerase II
  • DNA-Directed DNA Polymerase / isolation & purification
  • DNA-Directed DNA Polymerase / metabolism*
  • Dideoxynucleotides
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / enzymology*
  • Durapatite
  • Electrophoresis, Polyacrylamide Gel
  • Embryo, Nonmammalian / enzymology
  • Kinetics
  • Mammals
  • Molecular Weight
  • Poly dA-dT
  • Saccharomyces cerevisiae / enzymology
  • Templates, Genetic
  • Thymine Nucleotides / pharmacology

Substances

  • Dideoxynucleotides
  • Thymine Nucleotides
  • Poly dA-dT
  • Aphidicolin
  • Durapatite
  • DNA Polymerase II
  • DNA-Directed DNA Polymerase
  • 2',3'-dideoxythymidine triphosphate