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Mutations enabling displacement of tryptophan by 4-fluorotryptophan as a canonical amino acid of the genetic code.


ABSTRACT: The 20 canonical amino acids of the genetic code have been invariant over 3 billion years of biological evolution. Although various aminoacyl-tRNA synthetases can charge their cognate tRNAs with amino acid analogs, there has been no known displacement of any canonical amino acid from the code. Experimental departure from this universal protein alphabet comprising the canonical amino acids was first achieved in the mutants of the Bacillus subtilis QB928 strain, which after serial selection and mutagenesis led to the HR23 strain that could use 4-fluorotryptophan (4FTrp) but not canonical tryptophan (Trp) for propagation. To gain insight into this displacement of Trp from the genetic code by 4FTrp, genome sequencing was performed on LC33 (a precursor strain of HR23), HR23, and TR7 (a revertant of HR23 that regained the capacity to propagate on Trp). Compared with QB928, the negative regulator mtrB of Trp transport was found to be knocked out in LC33, HR23, and TR7, and sigma factor sigB was mutated in HR23 and TR7. Moreover, rpoBC encoding RNA polymerase subunits were mutated in three independent isolates of TR7 relative to HR23. Increased expression of sigB was also observed in HR23 and in TR7 growing under 4FTrp. These findings indicated that stabilization of the genetic code can be provided by just a small number of analog-sensitive proteins, forming an oligogenic barrier that safeguards the canonical amino acids throughout biological evolution.

SUBMITTER: Yu AC 

PROVIDER: S-EPMC3971595 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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Mutations enabling displacement of tryptophan by 4-fluorotryptophan as a canonical amino acid of the genetic code.

Yu Allen Chi-Shing AC   Yim Aldrin Kay-Yuen AK   Mat Wai-Kin WK   Tong Amy Hin-Yan AH   Lok Si S   Xue Hong H   Tsui Stephen Kwok-Wing SK   Wong J Tze-Fei JT   Chan Ting-Fung TF  

Genome biology and evolution 20140301 3


The 20 canonical amino acids of the genetic code have been invariant over 3 billion years of biological evolution. Although various aminoacyl-tRNA synthetases can charge their cognate tRNAs with amino acid analogs, there has been no known displacement of any canonical amino acid from the code. Experimental departure from this universal protein alphabet comprising the canonical amino acids was first achieved in the mutants of the Bacillus subtilis QB928 strain, which after serial selection and mu  ...[more]

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