Document Detail

Direct charging of tRNA(CUA) with pyrrolysine in vitro and in vivo.
MedLine Citation:
PMID:  15329732     Owner:  NLM     Status:  MEDLINE    
Pyrrolysine is the 22nd amino acid. An unresolved question has been how this atypical genetically encoded residue is inserted into proteins, because all previously described naturally occurring aminoacyl-tRNA synthetases are specific for one of the 20 universally distributed amino acids. Here we establish that synthetic L-pyrrolysine is attached as a free molecule to tRNA(CUA) by PylS, an archaeal class II aminoacyl-tRNA synthetase. PylS activates pyrrolysine with ATP and ligates pyrrolysine to tRNA(CUA) in vitro in reactions specific for pyrrolysine. The addition of pyrrolysine to Escherichia coli cells expressing pylT (encoding tRNA(CUA)) and pylS results in the translation of UAG in vivo as a sense codon. This is the first example from nature of direct aminoacylation of a tRNA with a non-canonical amino acid and shows that the genetic code of E. coli can be expanded to include UAG-directed pyrrolysine incorporation into proteins.
Sherry K Blight; Ross C Larue; Anirban Mahapatra; David G Longstaff; Edward Chang; Gang Zhao; Patrick T Kang; Kari B Green-Church; Michael K Chan; Joseph A Krzycki
Publication Detail:
Type:  Journal Article     Date:  2004-08-25
Journal Detail:
Title:  Nature     Volume:  431     ISSN:  1476-4687     ISO Abbreviation:  Nature     Publication Date:  2004 Sep 
Date Detail:
Created Date:  2004-09-16     Completed Date:  2004-10-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0410462     Medline TA:  Nature     Country:  England    
Other Details:
Languages:  eng     Pagination:  333-5     Citation Subset:  IM    
Department of Microbiology, 484 West 12th Avenue, The Ohio State University, Columbus, Ohio 43210, USA.
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MeSH Terms
Adenosine Triphosphate / metabolism
Amino Acyl-tRNA Synthetases / metabolism*
Anticodon / genetics
Archaea / enzymology
Archaeal Proteins
Cell-Free System
Codon / genetics
Diphosphates / metabolism
Escherichia coli / genetics
Genetic Code
Lysine / analogs & derivatives*,  metabolism*
Methyltransferases / chemistry,  genetics,  immunology,  metabolism
RNA, Transfer, Amino Acid-Specific / genetics,  metabolism*
Substrate Specificity
Suppression, Genetic / genetics
Reg. No./Substance:
0/Anticodon; 0/Archaeal Proteins; 0/Codon; 0/Diphosphates; 0/RNA, Transfer, Amino Acid-Specific; 0/pyrrolysine; 56-65-5/Adenosine Triphosphate; 56-87-1/Lysine; EC 2.1.1.-/Methyltransferases; EC 2.1.1.-/monomethylamine methyltransferase; EC 6.1.1.-/Amino Acyl-tRNA Synthetases
Comment In:
Nature. 2004 Sep 16;431(7006):257-8   [PMID:  15372017 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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