Open AccessRole of DNA · RNA Hybrids in Eukaryotes Purification of Two Ribonucleases H from Yeast Cells
Author(s) -
WYERS Françoise,
SENTENAC André,
FROMAGEOT Pierre
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10921.x
Subject(s) - ribonuclease , rna , dna , polynucleotide , transfer rna , ribonuclease iii , gel electrophoresis , polyacrylamide gel electrophoresis , biochemistry , chemistry , sodium dodecyl sulfate , chromatography , rnase p , lysis , amberlite , enzyme , gene , organic chemistry , adsorption , rna interference
Two ribonucleases H, which specifically degrade RNA in RNA‐DNA hybrid structures, have been extensively purified from extracts of Saccharomyces corevisiae. Ribonuclease H 1 can be obtained in high yield by a rapid purification procedure mainly involving ammonium sulfate fractionation and phosphocellulose chromatography. The enzyme preparation was shown to be homogeneous by acrylamide gel electrophoresis with sodium dodecylsulfate. Ribonuclease H 2 was also obtained in an essentially homogeneous form by phosphocellulose and Amberlite chromatography. About 60 mg of ribonuclease H 1 and 2 mg of ribonuclease H 2 were obtained from 100 g of wet cells. Using T7 DNA · RNA, hybrids, the specific activity of ribonuclease H 2 was approximately 10‐fold higher than that of ribonuclease H 1 .