PMID: 10971651Sep 6, 2000

The role of tightly bound ATP in Escherichia coli tRNA nucleotidyltransferase

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
Y TomariTakuya Ueda

Abstract

The CCA-adding enzyme [ATP(CTP): tRNA nucleotidyltransferase (EC. 2.7.7.25)] catalyses the addition of the conserved CCA sequence to the 3'-terminus of tRNAs. All CCA-adding enzymes are classified into the nucleotidyltransferase superfamily. In the absence of ATP, the Escherichia coli CCA-adding enzyme displays anomalous poly(C) polymerase activity. We show that CCA-adding enzyme over-expressed in E. coli exists in an ATP-bound form. The affinities of ATP and CTP towards the enzyme were estimated by several methods, and the dissociation constants for ATP and CTP were determined to be 6.3 and 188 microM, respectively. AMP-incorporation terminated the nucleotidyltransferase reaction, while in the absence of ATP, the enzyme continued poly(C) polymerization. In the case of a tRNA substrate with a mutation in the T-loop region, normal CC was added at a much slower rate compared with the wild-type, but anomalous poly(C) polymerization occurred at the same rate as in the wild-type. Based on the findings outlined above, we concluded that the E. coli CCA-adding enzyme possesses at least two distinct nucleotide binding sites, one responsible for ATP binding and the other(s) for CTP binding. The addition of ATP from the tight ATP binding ...Continue Reading

References

Jan 1, 1989·Methods in Enzymology·J F Milligan, O C Uhlenbeck
Feb 1, 1971·Archives of Biochemistry and Biophysics·A N Best, G D Novelli
Jan 28, 1994·Biochemical and Biophysical Research Communications·M P KalaposN Sarkar
Oct 29, 1998·The Journal of Biological Chemistry·D YueN Maizels

Citations

Jan 9, 2013·Nucleic Acids Research·Takaaki TaniguchiTsutomu Suzuki
Oct 23, 2004·Current Biology : CB·Alan M Weiner
Nov 9, 2001·The Journal of Biological Chemistry·Hyundae D ChoAlan M Weiner
Dec 1, 2005·The Journal of Biological Chemistry·Naoki ShigiK Watanabe
Aug 16, 2001·The Journal of Biological Chemistry·T NagaikeTakuya Ueda

Related Concepts

CCA-adding enzyme, human
Adenosine Triphosphate, Chromium Ammonium Salt
Molecular Sieve Chromatography
Mg CTP
SDS-PAGE
Alkalescens-Dispar Group
Poly C
Plasma Protein Binding Capacity
RNA Nucleotidyltransferases
Transfer RNA

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