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Nucleic Acids Res. Dec 25, 1992; 20(24): 6707–6712.
PMCID: PMC334590

Novel in-frame two codon translational hop during synthesis of bovine placental lactogen in a recombinant strain of Escherichia coli.


A recombinant Escherichia coli strain was constructed for the overexpression of bovine placental lactogen (bPL), using a bPL structural gene containing 9 of the rare arginine codons AGA and AGG. When high level bPL synthesis was induced in this strain, cell growth was inhibited and bPL accumulated to less than 10% of total cell protein. In addition, about 2% of the recombinant bPL produced from this strain exhibited an altered trypsin digestion pattern. Amino acid residues 74 through 109 normally produce 2 tryptic peptides, but the altered form of bPL lacked these two peptides and instead had a new peptide which was missing arginine residue 86 and one of the two flanking leucine residues. The codon for arginine residue 86 was AGG and the codons for the flanking leucine residues 85 and 87 were TTG. When 5 of the 9 AGA and AGG codons in the bPL structural gene were changed to more preferred arginine codons, cell growth was not inhibited and bPL accumulated to about 30% of total cell protein. When bPL was purified from this modified strain, which included changing the arginine codon at position 86 from AGG to CGT, none of the altered form of bPL was produced. These observations are consistent with a model in which translational pausing occurs at the arginine residue 86 AGG codon because the corresponding arginyl-tRNA species is reduced by the high level of bPL synthesis, and a translational hop occurs from the leucine residue 85 TTG codon to the leucine residue 87 TTG codon. This observation represents the first report of an error in protein synthesis due to an in-frame translational hop within an open reading frame.

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  • Bogosian G, Violand BN, Dorward-King EJ, Workman WE, Jung PE, Kane JF. Biosynthesis and incorporation into protein of norleucine by Escherichia coli. J Biol Chem. 1989 Jan 5;264(1):531–539. [PubMed]
  • Wada K, Aota S, Tsuchiya R, Ishibashi F, Gojobori T, Ikemura T. Codon usage tabulated from the GenBank genetic sequence data. Nucleic Acids Res. 1990 Apr 25;18 (Suppl):2367–2411. [PMC free article] [PubMed]
  • Obukowicz MG, Staten NR, Krivi GG. Enhanced heterologous gene expression in novel rpoH mutants of Escherichia coli. Appl Environ Microbiol. 1992 May;58(5):1511–1523. [PMC free article] [PubMed]
  • Hwang SO, Feldberg RS. Effect of inclusion body production on culture turbidity and cell dry weight in growing bacterial cultures. Biotechnol Prog. 1990 Jan-Feb;6(1):48–50. [PubMed]
  • O'Connor M, Gesteland RF, Atkins JF. tRNA hopping: enhancement by an expanded anticodon. EMBO J. 1989 Dec 20;8(13):4315–4323. [PMC free article] [PubMed]
  • Falahee MB, Weiss RB, O'Connor M, Doonan S, Gesteland RF, Atkins JF. Mutants of translational components that alter reading frame by two steps forward or one step back. J Biol Chem. 1988 Dec 5;263(34):18099–18103. [PubMed]
  • Weiss RB, Huang WM, Dunn DM. A nascent peptide is required for ribosomal bypass of the coding gap in bacteriophage T4 gene 60. Cell. 1990 Jul 13;62(1):117–126. [PubMed]
  • Huang WM, Ao SZ, Casjens S, Orlandi R, Zeikus R, Weiss R, Winge D, Fang M. A persistent untranslated sequence within bacteriophage T4 DNA topoisomerase gene 60. Science. 1988 Feb 26;239(4843):1005–1012. [PubMed]
  • Wong SC, Abdelal AT. Unorthodox expression of an enzyme: evidence for an untranslated region within carA from Pseudomonas aeruginosa. J Bacteriol. 1990 Feb;172(2):630–642. [PMC free article] [PubMed]
  • Seetharam R, Heeren RA, Wong EY, Braford SR, Klein BK, Aykent S, Kotts CE, Mathis KJ, Bishop BF, Jennings MJ, et al. Mistranslation in IGF-1 during over-expression of the protein in Escherichia coli using a synthetic gene containing low frequency codons. Biochem Biophys Res Commun. 1988 Aug 30;155(1):518–523. [PubMed]
  • Scorer CA, Carrier MJ, Rosenberger RF. Amino acid misincorporation during high-level expression of mouse epidermal growth factor in Escherichia coli. Nucleic Acids Res. 1991 Jul 11;19(13):3511–3516. [PMC free article] [PubMed]
  • Pedersen S. Escherichia coli ribosomes translate in vivo with variable rate. EMBO J. 1984 Dec 1;3(12):2895–2898. [PMC free article] [PubMed]
  • Lindsey DF, Mullin DA, Walker JR. Characterization of the cryptic lambdoid prophage DLP12 of Escherichia coli and overlap of the DLP12 integrase gene with the tRNA gene argU. J Bacteriol. 1989 Nov;171(11):6197–6205. [PMC free article] [PubMed]
  • Henson JM, Chu H, Irwin CA, Walker JR. Isolation and characterization of dnaX and dnaY temperature-sensitive mutants of Escherichia coli. Genetics. 1979 Aug;92(4):1041–1059. [PMC free article] [PubMed]
  • Garcia GM, Mar PK, Mullin DA, Walker JR, Prather NE. The E. coli dnaY gene encodes an arginine transfer RNA. Cell. 1986 May 9;45(3):453–459. [PubMed]
  • Atkins JF, Weiss RB, Gesteland RF. Ribosome gymnastics--degree of difficulty 9.5, style 10.0. Cell. 1990 Aug 10;62(3):413–423. [PubMed]
  • Weiss R, Gallant J. Mechanism of ribosome frameshifting during translation of the genetic code. Nature. 302(5907):389–393. [PubMed]
  • Weiss R, Lindsley D, Falahee B, Gallant J. On the mechanism of ribosomal frameshifting at hungry codons. J Mol Biol. 1988 Sep 20;203(2):403–410. [PubMed]
  • Spanjaard RA, van Duin J. Translation of the sequence AGG-AGG yields 50% ribosomal frameshift. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7967–7971. [PMC free article] [PubMed]
  • Robinson M, Lilley R, Little S, Emtage JS, Yarranton G, Stephens P, Millican A, Eaton M, Humphreys G. Codon usage can affect efficiency of translation of genes in Escherichia coli. Nucleic Acids Res. 1984 Sep 11;12(17):6663–6671. [PMC free article] [PubMed]
  • Bonekamp F, Andersen HD, Christensen T, Jensen KF. Codon-defined ribosomal pausing in Escherichia coli detected by using the pyrE attenuator to probe the coupling between transcription and translation. Nucleic Acids Res. 1985 Jun 11;13(11):4113–4123. [PMC free article] [PubMed]
  • Bonekamp F, Jensen KF. The AGG codon is translated slowly in E. coli even at very low expression levels. Nucleic Acids Res. 1988 Apr 11;16(7):3013–3024. [PMC free article] [PubMed]
  • Brinkmann U, Mattes RE, Buckel P. High-level expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene product. Gene. 1989 Dec 21;85(1):109–114. [PubMed]
  • Weiss RB, Dunn DM, Atkins JF, Gesteland RF. Slippery runs, shifty stops, backward steps, and forward hops: -2, -1, +1, +2, +5, and +6 ribosomal frameshifting. Cold Spring Harb Symp Quant Biol. 1987;52:687–693. [PubMed]
  • Weiss RB, Dunn DM, Atkins JF, Gesteland RF. Ribosomal frameshifting from -2 to +50 nucleotides. Prog Nucleic Acid Res Mol Biol. 1990;39:159–183. [PubMed]
  • VOGEL HJ, BONNER DM. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed]

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