Enlarging the amino acid set of Escherichia coli by infiltration of the valine coding pathway.
Aminoacyl transfer RNA (tRNA) synthetases establish the rules of the genetic code by catalyzing the aminoacylation of tRNAs. For some synthetases, accuracy depends critically on an editing function at a site distinct from the aminoacylation site. Mutants of Escherichia coli that incorrectly charge tRNA(Val) with cysteine were selected after random mutagenesis of the whole chromosome. All mutations obtained were located in the editing site of valyl-tRNA synthetase. More than 20% of the valine in cellular proteins from such an editing mutant organism could be replaced with the noncanonical aminobutyrate, sterically similar to cysteine. Thus, the editing function may have played a central role in restricting the genetic code to 20 amino acids. Disabling this editing function offers a powerful approach for diversifying the chemical composition of proteins and for emulating evolutionary stages of ambiguous translation.
Döring
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Evologic SA, 4 rue Pierre Fontaine, 91000 Evry, France.
Mootz
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Nangle
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Hendrickson
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TL
de Crécy-Lagard
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Schimmel
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Marlière
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P
D000483:Alleles; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000613:Aminobutyrates / Q000378:metabolism*; D001665:Binding Sites; D003062:Codon; D003545:Cysteine / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D005815:Genetic Code; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010641:Phenotype; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012366:RNA, Transfer, Val / Q000378:metabolism*; D013489:Suppression, Genetic; D013912:Threonine / Q000378:metabolism; D046249:Transfer RNA Aminoacylation; D014633:Valine / Q000378:metabolism; D014637:Valine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000613:Aminobutyrates; D003062:Codon; D012329:RNA, Bacterial; D012366:RNA, Transfer, Val; D013912:Threonine; D014637:Valine-tRNA Ligase; D014633:Valine; D003545:Cysteine