[Interaction of eukaryotic tyrosyl-tRNA-synthetase with high molecular weight RNA].
The affinity of eukaryotic tyrosyl-tRNA synthetases from bovine liver and from yeast for E. coli ribosomal RNA and synthetic polyribonucleotides has been studied by protein binding on the rRNA-Sepharose column and enzyme inhibition by high molecular weight RNAs. Tyrosyl-tRNA synthetase from bovine liver (Mr 2.59 kDa) was fully retained on the rRNA-Sepharose and eluted by buffer with 100 mM KCl. The functionally active modified form of bovine liver tyrosyl-tRNA synthetase obtained by endogenous limited proteolysis (Mr 2.38 kDa) partially maintains the affinity for rRNA and is eluted by 50 mM KCl. The highest rRNA-binding ability was revealed for yeast tyrosyl-tRNA synthetase eluted by 200 mM KCl. The E. coli tyrosyl-tRNA synthetase was not retained on rRNA-Sepharose. The aminoacylation activities of both bovine liver and yeast tyrosyl-tRNA synthetases were efficiently inhibited by rRNA and the inhibition was partially competitive in respect to tRNA(Tyr). At the same time the activities of proteolytically modified bovine tyrosyl-tRNA synthetase and E. coli tyrosyl-tRNA synthetase were not influenced by the addition of rRNA. Synthetic single- and double-stranded polyribonucleotides specifically inhibited the activity of bovine tyrosyl-tRNA synthetase to different extent. The inhibition degree of bovine liver tyrosyl-tRNA synthetase decreased in the order: poly (G) greater than poly (I) greater than poly (I).poly (C) greater than poly (G).poly (C) greater than poly (C) greater than poly (A). Poly (U) did not inhibit the activity of bovine liver tyrosyl-tRNA synthetase.
Kurochkin
I V
IV
Korneliuk
A I
AI
Matsuka
G Kh
GKh
D000818:Animals; D002384:Catalysis; D002417:Cattle; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000201:enzymology; D005057:Eukaryotic Cells / Q000201:enzymology*; D006868:Hydrolysis; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D012335:RNA, Ribosomal / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D016963:Thermus thermophilus / Q000201:enzymology; D014448:Tyrosine-tRNA Ligase / Q000378:metabolism*
D012335:RNA, Ribosomal; D014448:Tyrosine-tRNA Ligase
Vzaimodeĭstvie éukarioticheskoĭ tirozil-tRNK-sintetazy s vysokomolekuliarnymi RNK.