A kinetic analysis of the readdition of CMP or AMP into venom phosphodiesterase-digested transfer RNA by tRNAnucleotidyltransferase (EC 2.7.7.25) from Escherichia coli is presented.Five purified amino acid acceptor species and unfractionated tRNA were investigated.The K, value for the incorporation of AMP was lo-fold higher than the K,,, for CMP incorporation into tRNA from which two or three nucleotides had been removed.The K,,, values for ATP and CTP were similar for all tRNA tested.ATP inhibited CMP incorporation and CTP inhibited AMP incorporation, the K; values for the two nucleotides were the same, but larger than their K,,, values.The K, values for the tRNA substrates were about the same whether AMP or CMP incorporation was measured.The tRNA substrates from which one, two, or three nucleotides had been removed inhibited competitively the incorporation of AMP or CMP into tRNA.A smaller inhibition was observed with intact tRNA.
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Miller et al. (1971) studied this question.
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