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 language | English |
|---|---|
| Pages (from-to) | 50155-50159 |
| Number of pages | 5 |
| Journal | Journal of Biological Chemistry |
| Volume | 277 |
| Issue number | 51 |
| DOIs | |
| State | Published - Dec 20 2002 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
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 journal › Article
}
TY - JOUR
T1 - Mechanism of action of RNase T
T2 - I. Identification of residues required for catalysis, substrate binding, and dimerization
AU - Zuo, Yuhong
AU - Deutscher, Murray P
PY - 2002/12/20
Y1 - 2002/12/20
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0037147256&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037147256&partnerID=8YFLogxK
U2 - 10.1074/jbc.M207706200
DO - 10.1074/jbc.M207706200
M3 - Article
C2 - 12364334
AN - SCOPUS:0037147256
VL - 277
SP - 50155
EP - 50159
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 51
ER -