Evidence for a conserved relationship between an acceptor stem and a tRNA for aminoacylation.
The anticodon-independent aminoacylation of RNA hairpin helices that reconstruct tRNA acceptor stems has been demonstrated for at least 10 aminoacyl-tRNA synthetases. For Escherichia coli cysteine tRNA synthetase, the specificity of aminoacylation of the acceptor stem is determined by the U73 nucleotide adjacent to the amino acid attachment site. Because U73 is present in all known cysteine tRNAs, we investigated the ability of the E. coli cystein enzyme to aminoacylate a heterologous acceptor stem. We show here that a minihelixCys based on the acceptor-T psi C stem of yeast tRNACys is a substrate for the E. coli enzyme, and that aminoacylation of this minihelix is dependent on U73. Additionally, we identify two base pairs in the acceptor stem that quantitatively convert the E. coli acceptor stem to the yeast acceptor stem. The influence of U73 and these two base pairs is completely retained in the full-length tRNA. This suggests a conserved relationship between the acceptor stem alone and the acceptor stem in the context of a tRNA for aminoacylation with cysteine. However, the primary determinant in the species-specific aminoacylation of the E. coli and yeast cysteine tRNAs is a tertiary base pair at position 15:48 outside of the acceptor stem. Although E. coli tRNACys has an unusual G15:G48 tertiary base pair, yeast tRNACys has a more common G15:C48 that prevents efficient aminoacylation of yeast tRNACys by the E. coli enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)
Hou
Y M
YM
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Sterner
T
T
Bhalla
R
R
D000215:Acylation; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D001483:Base Sequence; D017124:Conserved Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005815:Genetic Code; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012350:RNA, Transfer, Cys / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity; D015003:Yeasts / Q000201:enzymology
D012350:RNA, Transfer, Cys; D000604:Amino Acyl-tRNA Synthetases; C019975:cysteinyl-tRNA synthetase