DNA-POLYMERASE-EPSILON FROM DROSOPHILA-MELANOGASTER

We identified a DNA polymerase species in Drosophila melanogaster embr yos, and purified it. This polymerase shared some common properties wi th 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 p rocessive in DNA synthesis, it cofractionates with 3'-5' exonuclease a ctivity, it is sensitive to aphidicolin and is resistance to ddTTP. Th e polymerase activity was inhibited in the immune-precipitation assay with anti-pol-epsilon antibodies, which were produced against a polype ptide coded on the cDNA of a putative Drosophila pol-epsilon we isolat ed previously. Using these antibodies, Western blot analysis revealed that this polymerase is a 250kDa polypeptide, which is the same size a s observed in mammals and yeast. These results indicate that Drosophil a produces the epsilon-class of DNA polymerase, and like mammals or ye ast, possesses the 5 typical classes of DNA polymerases (alpha to epsi lon) in its embryos. (C) 1997 Academic Press