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Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality

Pyrrolysine (Pyl), the 22(nd) natural amino acid, is genetically encoded by UAG and inserted into proteins by the unique suppressor tRNA(Pyl)1. The Methanosarcinaceae produce Pyl and express Pyl-containing methyltransferases that allow growth on methylamines2. Homologous methyltransferases and the P...

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Autores principales: Nozawa, Kayo, O’Donoghue, Patrick, Gundllapalli, Sarath, Araiso, Yuhei, Ishitani, Ryuichiro, Umehara, Takuya, Soll, Dieter, Nureki, Osamu
Formato: Texto
Lenguaje:English
Publicado: 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2648862/
https://www.ncbi.nlm.nih.gov/pubmed/19118381
http://dx.doi.org/10.1038/nature07611
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author Nozawa, Kayo
O’Donoghue, Patrick
Gundllapalli, Sarath
Araiso, Yuhei
Ishitani, Ryuichiro
Umehara, Takuya
Soll, Dieter
Nureki, Osamu
author_facet Nozawa, Kayo
O’Donoghue, Patrick
Gundllapalli, Sarath
Araiso, Yuhei
Ishitani, Ryuichiro
Umehara, Takuya
Soll, Dieter
Nureki, Osamu
author_sort Nozawa, Kayo
collection PubMed
description Pyrrolysine (Pyl), the 22(nd) natural amino acid, is genetically encoded by UAG and inserted into proteins by the unique suppressor tRNA(Pyl)1. The Methanosarcinaceae produce Pyl and express Pyl-containing methyltransferases that allow growth on methylamines2. Homologous methyltransferases and the Pyl biosynthetic and coding machinery are also found in two bacterial species1,3. Pyl coding is maintained by pyrrolysyl-tRNA synthetase (PylRS), which catalyzes the formation of Pyl-tRNA(Pyl)4,5. Pyl is not a recent addition to the genetic code. PylRS was already present in the last universal common ancestor6; it then persisted in organisms that utilize methylamines as energy sources. Recent protein engineering efforts added non-canonical amino acids to the genetic code7,8. This technology relies on the directed evolution of an ‘orthogonal’ tRNA synthetase:tRNA pair in which an engineered aminoacyl-tRNA synthetase (aaRS) specifically and exclusively acylates the orthogonal tRNA with a non-canonical amino acid. For Pyl the natural evolutionary process developed such a system some 3 billion years ago. When transformed into Escherichia coli, Methanosarcina barkeri PylRS and tRNA(Pyl) function as an orthogonal pair in vivo5,9. Here we demonstrate that Desulfitobacterium hafniense PylRS:tRNA(Pyl) is an orthogonal pair in vitro and in vivo, and present the crystal structure of this orthogonal pair. The ancient emergence of PylRS:tRNA(Pyl) allowed for the evolution of unique structural features in both the protein and the tRNA. These structural elements manifest an intricate, specialized aaRS:tRNA interaction surface highly distinct from those observed in any other known aaRS:tRNA complex; it is this general property that underlies the molecular basis of orthogonality.
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spelling pubmed-26488622009-08-26 Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality Nozawa, Kayo O’Donoghue, Patrick Gundllapalli, Sarath Araiso, Yuhei Ishitani, Ryuichiro Umehara, Takuya Soll, Dieter Nureki, Osamu Nature Article Pyrrolysine (Pyl), the 22(nd) natural amino acid, is genetically encoded by UAG and inserted into proteins by the unique suppressor tRNA(Pyl)1. The Methanosarcinaceae produce Pyl and express Pyl-containing methyltransferases that allow growth on methylamines2. Homologous methyltransferases and the Pyl biosynthetic and coding machinery are also found in two bacterial species1,3. Pyl coding is maintained by pyrrolysyl-tRNA synthetase (PylRS), which catalyzes the formation of Pyl-tRNA(Pyl)4,5. Pyl is not a recent addition to the genetic code. PylRS was already present in the last universal common ancestor6; it then persisted in organisms that utilize methylamines as energy sources. Recent protein engineering efforts added non-canonical amino acids to the genetic code7,8. This technology relies on the directed evolution of an ‘orthogonal’ tRNA synthetase:tRNA pair in which an engineered aminoacyl-tRNA synthetase (aaRS) specifically and exclusively acylates the orthogonal tRNA with a non-canonical amino acid. For Pyl the natural evolutionary process developed such a system some 3 billion years ago. When transformed into Escherichia coli, Methanosarcina barkeri PylRS and tRNA(Pyl) function as an orthogonal pair in vivo5,9. Here we demonstrate that Desulfitobacterium hafniense PylRS:tRNA(Pyl) is an orthogonal pair in vitro and in vivo, and present the crystal structure of this orthogonal pair. The ancient emergence of PylRS:tRNA(Pyl) allowed for the evolution of unique structural features in both the protein and the tRNA. These structural elements manifest an intricate, specialized aaRS:tRNA interaction surface highly distinct from those observed in any other known aaRS:tRNA complex; it is this general property that underlies the molecular basis of orthogonality. 2008-12-31 2009-02-26 /pmc/articles/PMC2648862/ /pubmed/19118381 http://dx.doi.org/10.1038/nature07611 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Nozawa, Kayo
O’Donoghue, Patrick
Gundllapalli, Sarath
Araiso, Yuhei
Ishitani, Ryuichiro
Umehara, Takuya
Soll, Dieter
Nureki, Osamu
Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title_full Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title_fullStr Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title_full_unstemmed Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title_short Pyrrolysyl-tRNA synthetase:tRNA(Pyl) structure reveals the molecular basis of orthogonality
title_sort pyrrolysyl-trna synthetase:trna(pyl) structure reveals the molecular basis of orthogonality
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2648862/
https://www.ncbi.nlm.nih.gov/pubmed/19118381
http://dx.doi.org/10.1038/nature07611
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