Molecular Cloning of the Gene Encoding Nuclear DNA Helicase II. A BOVINE HOMOLOGUE OF HUMAN RNA HELICASE A AND DROSOPHILA Mle PROTEIN (*)

Abstract

Nuclear DNA helicase II (NDH II) unwinds both DNA and RNA (Zhang, S., and Grosse, F.(1994) Biochemistry 33, 3906-3912). Here, we report on the molecular cloning and sequence determination of the complementary DNA (cDNA) coding for this DNA and RNA helicase. The full-length cDNA sequence was derived from overlapping clones that were detected by immunoscreening of a calf thymus cDNA library in bacteriophage gt11. This cDNA was 4,528 bases in length, which corresponded well with a 4.5-4.7-kilobase-long mRNA as detected by Northern blot analysis. The open reading frame of NDH II cDNA predicts a polypeptide of 1287 amino acids and a calculated molecular mass of 141,854 daltons. NDH II is related to a group of nucleic acid helicases from the DEAD/H box family II, with the signature motif DEIH in domain II. Two further proteins of this family, i.e. human RNA helicase A and Drosophila Maleless (Mle) protein, were found to be highly homologous to NDH II. With RNA helicase A, there was 91.5% identity and 95.5% similarity between the amino acid residues; with Mle protein, we observed a 50% identity and an 85% similarity. Antibodies against human RNA helicase A cross-reacted with NDH II, further supporting that NDH II is the bovine homologue of human RNA helicase A. Immunofluorescence studies revealed a mainly nuclear localization of NDH II. A role for NDH II in nuclear DNA and RNA metabolism is suggested.