A chimaeric glutamyl:glutaminyl-tRNA synthetase: implications for evolution
EPrints@IICB
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Title |
A chimaeric glutamyl:glutaminyl-tRNA synthetase: implications for evolution |
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Creator |
Saha, Rajesh
Dasgupta, Saumya Basu, Gautam Roy, Siddhartha |
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Subject |
Structural Biology & Bioinformatics
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Description |
aaRSs (aminoacyl-tRNA synthetases) are multi-domain proteins that have evolved by domain acquisition. The anti-codon binding domain was added to the more ancient catalytic domain during aaRS evolution. Unlike in eukaryotes, the anti-codon binding domains of GluRS (glutamyl-tRNA synthetase) and GlnRS (glutaminyl-tRNA synthetase) in bacteria are structurally distinct. This originates from the unique evolutionary history of GlnRSs. Starting from the catalytic domain, eukaryotic GluRS evolved by acquiring the archaea/eukaryote-specific anticodon binding domain after branching away from the eubacteria family. Subsequently, eukaryotic GlnRS evolved from GluRS by gene duplication and horizontally transferred to bacteria. In order to study the properties of the putative ancestral GluRS in eukaryotes, formed immediately after acquiring the anticodon binding domain, we have designed and constructed a chimaeric protein, cGluGlnRS, consisting of the catalytic domain, Ec GluRS (Escherichia coli GluRS), and the anticodon binding domain of EcGlnRS (E. coli GlnRS). In contrast to the isolated EcN-GluRS, cGluGlnRS showed detectable activity of glutamylation of E. coli tRNAglu and was capable of complementing an E. coli ts (temperature-sensitive)-GluRS strain at non-permissive temperatures. Both cGluGlnRS and EcNGluRS were found to bind E. coli tRNAglu with native EcGluRSlike affinity, suggesting that the anticodon-binding domain in cGluGlnRS enhances kcat for glutamylation. This was further confirmed from similar experiments with a chimaera between EcN-GluRS and the substrate-binding domain of EcDnaK (E. coli DnaK). We also show that an extended loop, present in the anticodon-binding domains of GlnRSs, is absent in archaeal GluRS, suggesting that the loop was a later addition, generating additional anti-codon discrimination capability in GlnRS as it evolved from GluRS in eukaryotes. |
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Date |
2009
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Type |
Article
PeerReviewed |
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Format |
application/pdf
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Identifier |
http://www.eprints.iicb.res.in/135/1/BIOCHEMICAL_JOURNAL%2C417%2C_449%2D455%2C2009[119].pdf
Saha, Rajesh and Dasgupta, Saumya and Basu, Gautam and Roy, Siddhartha (2009) A chimaeric glutamyl:glutaminyl-tRNA synthetase: implications for evolution. Biochem Journal, 417. pp. 449-455. |
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Relation |
http://dx.doi.org/10.1042/BJ20080747
http://www.eprints.iicb.res.in/135/ |
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