Open AccessPurification and some properties of a novel dsRNA degrading nuclease bound to rye germ ribosomes.
Author(s) -
Maria A. Siwecka
Publication year - 1997
Publication title -
acta biochimica polonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.452
H-Index - 78
eISSN - 1734-154X
pISSN - 0001-527X
DOI - 10.18388/abp.1997_4440
Subject(s) - nuclease , ribosome , sepharose , rna , affinity chromatography , polyacrylamide gel electrophoresis , biochemistry , enzyme , dna , molecular mass , micrococcal nuclease , chemistry , biology , rnase p , microbiology and biotechnology , gene , nucleosome , histone
A two-step procedure including affinity chromatography for purification of rye germ ribosomal nuclease that degrades double-stranded RNA from a virus of Penicillium chrysogenum and the poly(I).poly(C) complex was developed. The specific activity towards poly(I).poly(C) of the obtained nuclease preparations was 30 times as high as that of ribosomes. The recovery of activity was 3.4% when the Octyl-Sepharose column was used, and 2.0% in the case of the Phenyl-Sepharose column. On polyacrylamide/SDS gel electrophoresis the nuclease was resolved into two proteins of molecular mass 62 kDa and 57 kDa, respectively. 2-Mercaptoehanol and Mn2+ stimulated the activity of the purified enzyme. Glycerol (20%-50% concentration) stabilized enzyme. In addition to activity towards dsRNA and ssRNA the enzyme cleaves native and denatured DNA. It is suggested that this type of a nuclease takes part in regulation of the mRNA level in cytoplasm.