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Covalent complex of phenylalanyl-tRNA synthetase with 4-thiouridine-substituted tRNA(Phe) gene transcript retains
N A Moor1, A Favre, O I Lavrik
1Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Russian Academy of Sciences.
Abstract:
s4U-containing transcripts of tRNA(Phe) gene have been prepared by complete substitution of 16 U residues or by random incorporation of s4U residues followed by affinity electrophoresis isolation of s4U-monosubstituted tRNA transcripts. Both analogs have been cross-linked to Thermus thermophilus phenylalanyl-tRNA synthetase (PheRS) and the specificity of the cross-linking has been demonstrated. Functional activity of the covalent complex of PheRS with the s4U-monosubstituted transcript has been shown by aminoacylation of 60% of the enzyme-cross-linked tRNA. This is the first instance in which biological activity of aminoacyl-tRNA synthetase and cross-linked tRNA in a specific complex has been revealed.