title: Genetic-code evolution for protein synthesis with non-natural amino acids.
abstract: The genetic encoding of synthetic or "non-natural" amino acids promises to diversify the functions and structures of proteins. We applied rapid codon-reassignment for creating Escherichia coli strains unable to terminate translation at the UAG "stop" triplet, but efficiently decoding it as various tyrosine and lysine derivatives. This complete change in the UAG meaning enabled protein synthesis with these non-natural molecules at multiple defined sites, in addition to the 20 canonical amino acids. UAG was also redefined in the E. coli BL21 strain, suitable for the large-scale production of recombinant proteins, and its cell extract served the cell-free synthesis of an epigenetic protein, histone H4, fully acetylated at four specific lysine sites.
authors.affiliation: RIKEN Systems and Structural Biology Center, Tsurumi, Yokohama, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Mukai
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Yanagisawa
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Ohtake
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Wakamori
authors.forename: Jiro
authors.initials: J
authors.lastname: Adachi
authors.forename: Nobumasa
authors.initials: N
authors.lastname: Hino
authors.forename: Aya
authors.initials: A
authors.lastname: Sato
authors.forename: Takatsugu
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Akiko
authors.initials: A
authors.lastname: Hayashi
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.forename: Takashi
authors.initials: T
authors.lastname: Umehara
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Kensaku
authors.initials: K
authors.lastname: Sakamoto
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics*; D018388:Codon, Terminator / Q000235:genetics; D019020:Directed Molecular Evolution / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D019143:Evolution, Molecular; D055786:Gene Knockout Techniques; D005815:Genetic Code; D006657:Histones / Q000235:genetics / Q000378:metabolism; D010443:Peptide Chain Termination, Translational / Q000235:genetics; D010454:Peptide Termination Factors / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics*
chemical_list: D000596:Amino Acids; D018388:Codon, Terminator; D029968:Escherichia coli Proteins; D006657:Histones; D010454:Peptide Termination Factors; C489005:prfA protein, E coli