title: Enhanced phosphoserine insertion during Escherichia coli protein synthesis via partial UAG codon reassignment and release factor 1 deletion.
abstract: Genetically encoded phosphoserine incorporation programmed by the UAG codon was achieved by addition of engineered elongation factor and an archaeal aminoacyl-tRNA synthetase to the normal Escherichia coli translation machinery (Park et al., 2011) Science 333, 1151). However, protein yield suffers from expression of the orthogonal phosphoserine translation system and competition with release factor 1 (RF-1). In a strain lacking RF-1, phosphoserine phosphatase, and where seven UAG codons residing in essential genes were converted to UAA, phosphoserine incorporation into GFP and WNK4 was significantly elevated, but with an accompanying loss in cellular fitness and viability.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8144, USA.
authors.forename: Ilka U
authors.initials: IU
authors.lastname: Heinemann
authors.forename: Alexis J
authors.initials: AJ
authors.lastname: Rovner
authors.forename: Hans R
authors.initials: HR
authors.lastname: Aerni
authors.forename: Svetlana
authors.initials: S
authors.lastname: Rogulina
authors.forename: Laura
authors.initials: L
authors.lastname: Cheng
authors.forename: William
authors.initials: W
authors.lastname: Olds
authors.forename: Jonathan T
authors.initials: JT
authors.lastname: Fischer
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
authors.forename: Farren J
authors.initials: FJ
authors.lastname: Isaacs
authors.forename: Jesse
authors.initials: J
authors.lastname: Rinehart
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018388:Codon, Terminator / Q000235:genetics*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics*; D017353:Gene Deletion; D016680:Genome, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D010454:Peptide Termination Factors / Q000172:deficiency* / Q000235:genetics*; D010641:Phenotype; D010768:Phosphoserine / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000235:genetics*; D020543:Proteome / Q000235:genetics
chemical_list: D018388:Codon, Terminator; D029968:Escherichia coli Proteins; D010454:Peptide Termination Factors; D020543:Proteome; C489005:prfA protein, E coli; D010768:Phosphoserine