An engineered class I transfer RNA with a class II tertiary fold.
Structure-based engineering of the tertiary fold of Escherichia coli tRNA(Gln)2 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 tRNA(Ser). 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.
Nissan
T A
TA
Department of Chemistry, University of California at Santa Barbara 93106-9510, USA.
Oliphant
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B
Perona
J J
JJ
D000215:Acylation; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D012329:RNA, Bacterial / Q000737:chemistry; D012351:RNA, Transfer, Gln / Q000737:chemistry*; D012362:RNA, Transfer, Ser / Q000737:chemistry; D016415:Sequence Alignment; D013696:Temperature; D016963:Thermus thermophilus / Q000235:genetics
D012329:RNA, Bacterial; D012351:RNA, Transfer, Gln; D012362:RNA, Transfer, Ser