Intracellular localization of DNA polymerase alpha

Abstract

Immunoglobulin (IgG) and the F(ab')2 fragment of IgG were prepared from serum of a rabbit immunized with purified calf thymus DNA polymerase alpha (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7). An indirect immunofluorescent method based on these reagents was used to detect the intracellular localization of DNA polymerase alpha in primary fetal bovine fibroblasts. The results show that the bulk of DNA polymerase alpha is located in the perinuclear region of the cytoplasm. Immunofluorescent staining of cytoplast and Ficoll-Paque gradient-purified karyoplast fragments resulting from cytochalasin enucleation show the presence of DNA polymerase alpha in cytoplasts and the virtual absence of the enzyme in the nucleus of the karyoplast itself. The implication of this unusual intracellular location for DNA polymerase alpha is discussed.