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Reference: Favorova OO, et al. (1981) Partial digestion of tRNA--aminoacyl-tRNA synthetase complexes with cobra venom ribonuclease. Biochemistry 20(4):1006-11

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Abstract


Transfer RNA molecules have been labeled with 32P at the 5' or 3' end and digested with cobra venom ribonuclease, which preferentially cuts double-stranded regions. The products of yeast tRNAPhe and tRNAVal were analyzed by high-resolution gel electrohporesis. In the free state, these tRNAs were cut predominantly in the acceptor and anticodon stems. Minor cuts occurred in the T psi stem in tRNAVal. The topography of zones interacting with their cognate synthetases was studied by determining the tRNA regions shielded by protein. Nearly 100% protection was found in the anticodon and acceptor stem of tRNAVal, while in tRNAPhe only the stem of the anticodon was protected. Noncognate interactions between tRNAPhe and tryptophanyl-tRNA synthetase from beef pancreas were examined. The beef enzyme did not protect tRNAPhe despite the fact that efficient misaminoacylation occurred. The pattern of shielding obtained for each tRNA-synthetase complex was compared with the results of direct ultraviolet cross-linking experiments with these complexes.

Reference Type
Journal Article
Authors
Favorova OO, Fasiolo F, Keith G, Vassilenko SK, Ebel JP
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