Interaction of ribonuclease H from Drosophila melanogaster embryos with DNA polymerase-primase.

An RNase H was purified 2,500-fold to near homogeneity from early embryos of Drosophila melanogaster. The purified enzyme has an approximate molecular weight of 180,000 and appears to consist of two 49,000- and two 39,000-dalton polypeptides. The enzyme specifically hydrolyzes RNA.DNA hybrids and releases oligoribonucleotides ranging in size from 2-9 residues. The RNase H can also remove RNA primers that are synthesized and subsequently elongated by the Drosophila polymerase-primase. Preincubation of the RNase H from D. melanogaster embryos with the homologous DNA polymerase-primase results in an increased rate of DNA synthesis. The DNA chains synthesized under these conditions are shorter than those synthesized in the absence of the RNase H, and the rate of primer synthesis is increased significantly. These findings suggest that the RNase H forms a complex with the polymerase-primase, increasing its recycling capacity and thereby increasing the frequency of chain initiation.