PMID:12364334
| Citation |
Zuo, Y and Deutscher, MP (2002) Mechanism of action of RNase T. I. Identification of residues required for catalysis, substrate binding, and dimerization. J. Biol. Chem. 277:50155-9 |
|---|---|
| Abstract |
Escherichia coli RNase T, an RNA-processing enzyme and a member of the DEDD exonuclease superfamily, was examined using sequence analysis and site-directed mutagenesis. Like other DEDD exonucleases, RNase T was found to contain three conserved Exo motifs that included four invariant acidic residues. Mutagenesis of these motifs revealed that they are essential for RNase T activity, indicating that they probably form the RNase T catalytic center in a manner similar to that found in other DEDD exonucleases. We also identified by sequence analysis three short, but highly conserved, sequence segments rich in positively charged residues. Site-directed mutagenesis of these regions indicated that they are involved in substrate binding. Additional analysis revealed that residues within the C-terminal region of RNase T are essential for RNase T dimerization and, consequently, for RNase T activity. These data define the domains necessary for RNase T action, and together with information in the accompanying article, have led to the formulation of a detailed model for the structure and mechanism of action of RNase T. |
| Links |
PubMed Online version:10.1074/jbc.M207706200 |
| Keywords |
Amino Acid Motifs; Amino Acid Sequence; Catalysis; Chromatography, Gel; Dimerization; Escherichia coli/enzymology; Exoribonucleases/chemistry; Exoribonucleases/genetics; Exoribonucleases/metabolism; Kinetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Plasmids/metabolism; Protein Binding; Protein Structure, Tertiary; RNA, Transfer/metabolism; Sequence Homology, Amino Acid; Temperature |
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