DNA polymerase epsilon from Drosophila melanogaster.
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.
Aoyagi
N
N
Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, Noda-shi, Japan.
Oshige
M
M
Hirose
F
F
Kuroda
K
K
Matsukage
A
A
Sakaguchi
K
K
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D015153:Blotting, Western; D002845:Chromatography; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D054306:Dideoxynucleotides; D004331:Drosophila melanogaster / Q000196:embryology / Q000201:enzymology*; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004625:Embryo, Nonmammalian / Q000201:enzymology; D007700:Kinetics; D008322:Mammals; D008970:Molecular Weight; D011067:Poly dA-dT; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000494:pharmacology
D054306:Dideoxynucleotides; D013942:Thymine Nucleotides; D011067:Poly dA-dT; D016590:Aphidicolin; D017886:Durapatite; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase; C017216:2',3'-dideoxythymidine triphosphate