PMID:10889033

Citation	Pleiss, JA, Wolfson, AD and Uhlenbeck, OC (2000) Mapping contacts between Escherichia coli alanyl tRNA synthetase and 2' hydroxyls using a complete tRNA molecule. Biochemistry 39:8250-8
Abstract	A dual-specific derivative of yeast tRNA(Phe) is described whose features facilitate structure-function studies of tRNAs. This tRNA has been made in three different bimolecular forms that allow modifications to be easily introduced into any position within the molecule. A set of deoxynucleotide substituted versions of this tRNA has been created and used to examine contacts between tRNA and Escherichia coli alanyl-tRNA synthetase, an enzyme previously shown to interact with 2'-hydroxyls in the acceptor stem of the tRNA. Because the present experiments used a full-length tRNA, several contacts were identified that had not been previously found using microhelix substrates. Contacts at similar sites in the T-loop are seen in the cocrystal structure of tRNA(Ser) and Thermus thermophilus seryl-tRNA synthetase.
Links	PubMed
Keywords	Acylation; Alanine-tRNA Ligase/chemistry; Alanine-tRNA Ligase/metabolism; Base Sequence; Escherichia coli/enzymology; Escherichia coli/genetics; Hydroxyl Radical/metabolism; Kinetics; Models, Molecular; Molecular Sequence Data; Nucleic Acid Conformation; RNA, Bacterial/chemistry; RNA, Bacterial/metabolism; RNA, Transfer/chemistry; RNA, Transfer/metabolism
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