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Purification of Saccharomyces cerevisiae RNase H(70) and identification of the corresponding gene
Author(s) -
Frank Peter,
Braunshofer-Reiter Christa,
Karwan Anneliese,
Grimm Rudolf,
Wintersberger Ulrike
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00512-8
Subject(s) - saccharomyces cerevisiae , rnase p , gene , chemistry , identification (biology) , biochemistry , genetics , computational biology , microbiology and biotechnology , biology , rna , botany
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 62 849. 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.