Species-specific differences in amino acid editing by class II prolyl-tRNA synthetase.
Aminoacyl-tRNA synthetases are a family of enzymes responsible for ensuring the accuracy of the genetic code by specifically attaching a particular amino acid to their cognate tRNA substrates. Through primary sequence alignments, prolyl-tRNA synthetases (ProRSs) have been divided into two phylogenetically divergent groups. We have been interested in understanding whether the unusual evolutionary pattern of ProRSs corresponds to functional differences as well. Previously, we showed that some features of tRNA recognition and aminoacylation are indeed group-specific. Here, we examine the species-specific differences in another enzymatic activity, namely amino acid editing. Proofreading or editing provides a mechanism by which incorrectly activated amino acids are hydrolyzed and thus prevented from misincorporation into proteins. "Prokaryotic-like" Escherichia coli ProRS has recently been shown to be capable of misactivating alanine and possesses both pretransfer and post-transfer hydrolytic editing activity against this noncognate amino acid. We now find that two ProRSs belonging to the "eukaryotic-like" group exhibit differences in their hydrolytic editing activity. Whereas ProRS from Methanococcus jannaschii is similar to E. coli in its ability to hydrolyze misactivated alanine via both pretransfer and post-transfer editing pathways, human ProRS lacks these activities. These results have implications for the selection or design of antibiotics that specifically target the editing active site of the prokaryotic-like group of ProRSs.
Beuning
P J
PJ
Department of Chemistry, University of Minnesota, Minneapolis 55455, USA.
Musier-Forsyth
K
K
D000409:Alanine / Q000494:pharmacology; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D006801:Humans; D017017:Methanococcus / Q000201:enzymology*; D008969:Molecular Sequence Data; D011392:Proline / Q000494:pharmacology; D017393:RNA Editing; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D013045:Species Specificity
D000596:Amino Acids; D012346:RNA, Transfer, Amino Acyl; D011392:Proline; D000604:Amino Acyl-tRNA Synthetases; D000409:Alanine