Hydrolytic Editing by IleRS and ValRS
A noncognate dNTP is also misaligned with respect to catalytic
residues; formation of the phosphodiester bond proceeds 1000-
fold more slowly than with the cognate dNTP. Together, the
slow chemical step and reversible conformational change
greatly enhance dissociation of the noncognate dNTP. It was
further suggested that this utilization of induced fit (during the
mismatch recognition) to slow catalysis and promote noncog-
nate substrate release may be widely used to increase selectivity
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(
51). However, our data for class I AARS clearly show that cog-
nate and noncognate amino acids are each transferred to tRNA
with similar rates under saturating conditions. Thus, misalign-
1
ment of reactive groups in the chemical step is not a component 16. Bishop, A. C., Nomanbhoy, T. K., and Schimmel, P. (2002) Proc. Natl.
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of amino acid selectivity by ValRS and IleRS.
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Absence of specificity at the tRNA transfer step appears not
to be confined to class I AARS, because the class II ThrRS trans-
Thr
fers noncognate serine and cognate threonine to tRNA with
1
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ilar measurements on other enzymes.
The double-sieve model for editing AARS proposes that the
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this pre-transfer editing sieve. This could have provided an evo-
lutionary driving force for the addition of the separate post-
transfer editing domain to a primordial editing tRNA synthe-
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Acknowledgments—We thank Ya-Ming Hou (Thomas Jefferson Uni-
versity, Philadelphia) for the IleRS and ValRS overexpression plas-
3
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tical Course: Transient Kinetic Methods Applied to Biological Mac-
romolecules (UK, 2008), and this training helped her in developing
transient kinetic methods at the University of Zagreb, Croatia.
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