Structure-based engineering of the tertiary fold
of Escherichia coli tRNA2Gln
has enabled conversion of this transfer RNA to a class II
structure while retaining recognition properties of a
class I glutamine tRNA. The new tRNA possesses the
20-nt variable stem-loop of Thermus thermophilus
tRNASer. Enlargement of the D-loop appears essential
to maintaining a stable tertiary structure in this species,
while rearrangement of a base triple in the augmented D-stem
is critical for efficient glutaminylation. These data provide
new insight into structural determinants distinguishing
the class I and class II tRNA folds, and demonstrate a
marked sensitivity of glutaminyl-tRNA synthetase to alteration
of tRNA tertiary structure.