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Recognition in vitro of the suppressor tRNA Pyl by the class II‐like Pyrrolsyl‐tRNA Synthetase
Author(s) -
Herring Stephanie Chiyoko,
Li Darrick,
Ambrogelly Alexandre,
Polycarpo Carla,
Soll Dieter
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a503-b
Subject(s) - transfer rna , amino acid , enzyme , biochemistry , aminoacyl trna synthetase , biology , chemistry , rna , gene
Pyrrolysine (Pyl) is the 22 nd amino acid cotranslationally inserted in response to a UAG codon. Pyrrolysyl‐tRNA synthetase (PylRS) is specific for attaching pyrrolysine to its cognate tRNA species that recognizes UAG (tRNA Pyl ). Analysis of the known genomes reveals that this enzyme:tRNA pair is present in the Methanosarcinaceae , Methanococcoides burtonii , and in the bacteria Desulfitobacterium hafniense and Desulfitobacterium frappieri . We wanted to compare the cross‐species specificity of the PylRS:tRNA Pyl interaction by studying tRNA charging by the different enzymes, as well as the binding affinities and footprints of enzyme:tRNA complexes. Our results show that while there are minor variations, all PylRS enzymes are able to recognize tRNA Pyl . Charging of tRNA Lys with lysine was not catalyzed by any PylRS. Interestingly, D. hafniense PylRS has an approximately 100 amino acid truncation at the amino terminus, yet it also recognizes tRNA Pyl . This suggests the amino terminus may not be essential for amino acid charging. (This work was supported by grants from NIGMS, National Institutes of Health, and the U. S. Department of Energy).