How a CCA Sequence Protects Mature tRNAs and tRNA Precursors from Action of the Processing Enzyme RNase BN/RNase Z |
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Authors: | Tanmay Dutta Arun Malhotra Murray P. Deutscher |
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Affiliation: | From the Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, Florida 33136 |
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Abstract: | In many organisms, 3′ maturation of tRNAs is catalyzed by the endoribonuclease, RNase BN/RNase Z, which cleaves after the discriminator nucleotide to generate a substrate for addition of the universal CCA sequence. However, tRNAs or tRNA precursors that already contain a CCA sequence are not cleaved, thereby avoiding a futile cycle of removal and readdition of these essential residues. We show here that the adjacent C residues of the CCA sequence and an Arg residue within a highly conserved sequence motif in the channel leading to the RNase catalytic site are both required for the protective effect of the CCA sequence. When both of these determinants are present, CCA-containing RNAs in the channel are unable to move into the catalytic site; however, substitution of either of the C residues by A or U or mutation of Arg274 to Ala allows RNA movement and catalysis to proceed. These data define a novel mechanism for how tRNAs are protected against the promiscuous action of a processing enzyme. |
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Keywords: | Enzyme Structure Nucleic Acid Chemistry Nucleic Acid Enzymology Phosphodiesterases Protein-Nucleic Acid Interaction RNA Catalysis RNA Processing RNA-Protein Interaction Structural Biology Transfer RNA (tRNA) |
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