title: Purification of Saccharomyces cerevisiae RNase H(70) and identification of the corresponding gene.
abstract: We purified Saccharomyces cerevisiae RNase H(70) to homogeneity, using an optimized chromatographic purification procedure. Renaturation gel assay assigned RNase H activity to a 70 kDa polypeptide. Sequencing of tryptic peptides identified the open reading frame YGR276c on chromosome VII of the S. cerevisiae genome as the corresponding gene, which encodes a putative polypeptide of molecular mass of 62849. We therefore renamed this gene RNH70. Immunofluorescence microscopy using a RNH70-EGFP fusion construct indicates nuclear localization of RNase H(70). Deletion of RNH70 from the yeast genome did not result in any serious phenotype under the conditions tested. Homology searches revealed striking similarity with a number of eukaryotic proteins and open reading frames, among them the chimpanzee GOR protein, a homolog of a human autoimmune antigen, found to elicit autoimmune response in patients infected with hepatitis C virus.
authors.affiliation: Department of Molecular Genetics, Institute of Tumor Biology and Cancer Research, University of Vienna, Austria.
authors.forename: P
authors.initials: P
authors.lastname: Frank
authors.forename: C
authors.initials: C
authors.lastname: Braunshofer-Reiter
authors.forename: A
authors.initials: A
authors.lastname: Karwan
authors.forename: R
authors.initials: R
authors.lastname: Grimm
authors.forename: U
authors.initials: U
authors.lastname: Wintersberger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus; D005057:Eukaryotic Cells; D005800:Genes, Fungal; D016681:Genome, Fungal; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016914:Ribonuclease H / Q000235:genetics / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D016914:Ribonuclease H