Recognition of tRNA(Gly) by three widely diverged glycyl-tRNA synthetases.
Glycyl-tRNA synthetase (GlyRS) is an unusual aminoacyl-tRNA synthetase because it varies in its quarternary structure between organisms; Escherichia coli GlyRS is an alpha2beta2 tetramer, whereas those of Thermus thermophilus and yeast are alpha2 dimers. In contrast, the tRNA(Gly) sequence is virtually identical in E. coli and T. thermophilus but very different in yeast. In this study, we examined the molecular recognition of tRNA(Gly) by three widely diverged GlyRSs using in vitro tRNA transcripts. Mutation studies showed that the discriminator base at position 73, the second base-pair, C2 x G71, in the acceptor stem, and the anticodon nucleotides, C35 and C36, contribute to the specific aminoacylation of all three GlyRSs, the discriminator base differing between prokaryotes (U73) and eukaryotes (A73). However, we found differences between yeast and two bacteria around the second base-pair in the acceptor stem. The first base-pair, G1 x C72, is important for glycylation in E. coli and T. thermophilus, whereas the third base-pair, G3 x C70, is important for glycylation in yeast. These findings indicate that despite such large differences of the two prokaryotic GlyRSs, tRNA(Gly) identity has been essentially conserved in prokaryotes, and that there are also differences in the acceptor stem recognition between prokaryotes and yeast. The clear separation between prokaryotes and yeast is retained in the identity element location, whereas the apparent diversity of the two prokaryotic enzymes does not reflect on the tRNA recognition.
Nameki
N
N
Department of Biology, Faculty of Science, Hirosaki University, Japan.
Tamura
K
K
Asahara
H
H
Hasegawa
T
T
D000215:Acylation; D000926:Anticodon / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology*; D006032:Glycine-tRNA Ligase / Q000737:chemistry*; D007700:Kinetics; D008969:Molecular Sequence Data; D017354:Point Mutation; D012353:RNA, Transfer, Gly / Q000737:chemistry* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D016963:Thermus thermophilus / Q000201:enzymology*
D000926:Anticodon; D012353:RNA, Transfer, Gly; D006032:Glycine-tRNA Ligase