Ja. Pleiss et al., Mapping contacts between Escherichia coli alanyl tRNA synthetase and 2 ' hydroxyls using a complete tRNA molecule, BIOCHEM, 39(28), 2000, pp. 8250-8258
A dual-specific derivative of yeast tRNA(Phe) is described whose features f
acilitate structure-function studies of tRNAs. This tRNA has been made in t
hree different bimolecular forms that allow modifications to be easily intr
oduced into any position within the molecule. A set of deoxynucleotide subs
tituted versions of this tRNA has been created and used to examine contacts
between tRNA and Escherichia coli alanyl-tRNA synthetase, an enzyme previo
usly shown to interact with 2'-hydroxyls in the acceptor stem of the tRNA.
Because the present experiments used a full-length tRNA, several contacts w
ere identified that had not been previously found using microhelix substrat
es. Contacts at similar sites in the T-loop are seen in the cocrystal struc
ture of tRNA(Ser) and Thermus thermophilus seryl-tRNA synthetase.