Mechanism of action of RNase T: I. Identification of residues required for catalysis, substrate binding, and dimerization

Yuhong Zuo, Murray P Deutscher

Research output: Contribution to journalArticle

19 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)50155-50159
Number of pages5
JournalJournal of Biological Chemistry
Volume277
Issue number51
DOIs
StatePublished - Dec 20 2002

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Pancreatic Ribonuclease
Dimerization
Catalysis
Substrates
Exonucleases
Mutagenesis
Site-Directed Mutagenesis
Sequence Analysis
exoribonuclease T
Conserved Sequence
Escherichia coli
RNA

ASJC Scopus subject areas

  • Biochemistry

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Mechanism of action of RNase T : I. Identification of residues required for catalysis, substrate binding, and dimerization. / Zuo, Yuhong; Deutscher, Murray P.

In: Journal of Biological Chemistry, Vol. 277, No. 51, 20.12.2002, p. 50155-50159.

Research output: Contribution to journalArticle

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