title: Suppression of amber codons in Caulobacter crescentus by the orthogonal Escherichia coli histidyl-tRNA synthetase/tRNAHis pair.
abstract: While translational read-through of stop codons by suppressor tRNAs is common in many bacteria, archaea and eukaryotes, this phenomenon has not yet been observed in the α-proteobacterium Caulobacter crescentus. Based on a previous report that C. crescentus and Escherichia coli tRNA(His) have distinctive identity elements, we constructed E. coli tRNA(His) CUA, a UAG suppressor tRNA for C. crescentus. By examining the expression of three UAG codon- containing reporter genes (encoding a β-lactamase, the fluorescent mCherry protein, or the C. crescentus xylonate dehydratase), we demonstrated that the E. coli histidyl-tRNA synthetase/tRNA(His) CUA pair enables in vivo UAG suppression in C. crescentus. E. coli histidyl-tRNA synthetase (HisRS) or tRNA(His) CUA alone did not achieve suppression; this indicates that the E. coli HisRS/tRNA(His) CUA pair is orthogonal in C. crescentus. These results illustrate that UAG suppression can be achieved in C. crescentus with an orthogonal aminoacyl-tRNA synthetase/suppressor tRNA pair.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, United States of America.
authors.forename: Jae-hyeong
authors.initials: JH
authors.lastname: Ko
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut, United States of America.
authors.forename: Paula Montero
authors.initials: PM
authors.lastname: Llopis
authors.affiliation: Howard Hughes Medical Institute, Yale University, New Haven, Connecticut, United States of America.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Heinritz
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut, United States of America ; Howard Hughes Medical Institute, Yale University, New Haven, Connecticut, United States of America ; Department of Microbial Pathogenesis, Yale School of Medicine, New Haven, Connecticut, United States of America.
authors.forename: Christine
authors.initials: C
authors.lastname: Jacobs-Wagner
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, United States of America ; Department of Chemistry, Yale University, New Haven, Connecticut, United States of America.
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
mesh_terms: D000668:Ampicillin Resistance / Q000235:genetics; D016935:Caulobacter crescentus / Q000235:genetics* / Q000378:metabolism*; D018388:Codon, Terminator; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D006643:Histidine-tRNA Ligase / Q000378:metabolism*; D012354:RNA, Transfer, His / Q000235:genetics* / Q000378:metabolism
chemical_list: D018388:Codon, Terminator; D012354:RNA, Transfer, His; D006643:Histidine-tRNA Ligase