N. Aoyagi et al., DROSOPHILA DNA-POLYMERASE-DELTA - PURIFICATION AND CHARACTERIZATION, The Journal of biological chemistry, 269(8), 1994, pp. 6045-6050
A DNA polymerase with properties similar to mammalian polymerase delta
has been isolated to near homogeneity from early embryos of Drosophil
a melanogaster. A combination of exclusion chromatography and sodium d
odecyl sulfate-polyacrylamide gel electrophoresis indicates that this
enzyme has a total molecular mass of 185 kDa and is composed of 138- a
nd 47-kDa polypeptides. Its isoelectric point is 6.8. This polymerase
activity is strongly inhibited by N-ethylmaleimide, aphidicolin, and h
igh KCl concentration but is relatively insensitive to 2',3' dideoxyth
ymidine 5'-triphosphate. There was no reaction in an immunological tes
t using monoclonal antibody against Drosophila DNA polymerase alpha. I
n a final purification step, this polymerase activity was accompanied
by 3' --> 5' exonuclease activity as expected proofreading activity. T
his polymerase activity is remarkably stimulated by mouse proliferatin
g cell nuclear antigen, which is structurally and immunologically very
similar to a Drosophila counterpart. These properties clearly indicat
e this enzyme belongs to the category of DNA polymerase delta.