Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. 1985 Sep; 82(17): 5656–5660.
PMCID: PMC390610

Translational efficiency of the Escherichia coli adenylate cyclase gene: mutating the UUG initiation codon to GUG or AUG results in increased gene expression.


Roy et al. [Roy, A., Haziza, C. & Danchin, A. (1983) EMBO J. 2, 791-797] established that translation of Escherichia coli adenylate cyclase initiates at a UUG codon, and they suggested this might decrease the efficiency of translation. We investigated the effect of varying the initiation codon on the expression of the adenylate cyclase (cya) gene. Using oligonucleotide-directed mutagenesis, we changed the UUG initiation codon to GUG and the more common initiator AUG and assayed for cya gene expression in a number of ways. First, the GUG initiation codon, in place of UUG, doubled cya expression when cya was expressed from the dual cya P1/P2 promoters. The corresponding AUG codon construct was nonviable. Second, when the cya gene was placed under the transcriptional control of the thermoinducible phage lambda PL promoter, the relative amounts of cya gene product were 1:2:6 for the UUG, GUG, and AUG initiation codons, respectively. Finally, the cya P2 promoter, Shine-Dalgarno sequence, and the DNA corresponding to the first 86 codons of cya were fused to DNA encoding the E. coli galactokinase gene beginning at the second codon. The relative amounts of the fusion polypeptides, which had galactokinase activity, were 1:2:3 for the UUG, GUG, and AUG initiation codons, respectively. These results demonstrate that the cya UUG initiation codon limits cya expression at the level of translation.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Gold L, Pribnow D, Schneider T, Shinedling S, Singer BS, Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. [PubMed]
  • Kozak M. Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. Microbiol Rev. 1983 Mar;47(1):1–45. [PMC free article] [PubMed]
  • Clark BF, Marcker KA. The role of N-formyl-methionyl-sRNA in protein biosynthesis. J Mol Biol. 1966 Jun;17(2):394–406. [PubMed]
  • Roy A, Haziza C, Danchin A. Regulation of adenylate cyclase synthesis in Escherichia coli: nucleotide sequence of the control region. EMBO J. 1983;2(5):791–797. [PMC free article] [PubMed]
  • Danchin A, Guiso N, Roy A, Ullmann A. Identification of the Escherichia coli cya gene product as authentic adenylate cyclase. J Mol Biol. 1984 May 25;175(3):403–408. [PubMed]
  • Aiba H, Mori K, Tanaka M, Ooi T, Roy A, Danchin A. The complete nucleotide sequence of the adenylate cyclase gene of Escherichia coli. Nucleic Acids Res. 1984 Dec 21;12(24):9427–9440. [PMC free article] [PubMed]
  • Yang JK, Epstein W. Purification and characterization of adenylate cyclase from Escherichia coli K12. J Biol Chem. 1983 Mar 25;258(6):3750–3758. [PubMed]
  • Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. [PubMed]
  • Roy A, Danchin A. The cya locus of Escherichia coli K12: organization and gene products. Mol Gen Genet. 1982;188(3):465–471. [PubMed]
  • Rao RN. Construction and properties of plasmid pKC30, a pBR322 derivative containing the pL-N region of phage lambda. Gene. 1984 Nov;31(1-3):247–250. [PubMed]
  • Schümperli D, McKenney K, Sobieski DA, Rosenberg M. Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon. Cell. 1982 Oct;30(3):865–871. [PubMed]
  • Messing J, Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Sanger F, Coulson AR, Barrell BG, Smith AJ, Roe BA. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. [PubMed]
  • Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. [PubMed]
  • Winter G, Fersht AR, Wilkinson AJ, Zoller M, Smith M. Redesigning enzyme structure by site-directed mutagenesis: tyrosyl tRNA synthetase and ATP binding. Nature. 1982 Oct 21;299(5885):756–758. [PubMed]
  • VOGEL HJ, BONNER DM. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed]
  • Rosenberg M, Ho YS, Shatzman A. The use of pKc30 and its derivatives for controlled expression of genes. Methods Enzymol. 1983;101:123–138. [PubMed]
  • Liberman E, Reddy P, Gazdar C, Peterkofsky A. The Escherichia coli adenylate cyclase complex. Stimulation by potassium and phosphate. J Biol Chem. 1985 Apr 10;260(7):4075–4081. [PubMed]
  • McKenney K, Shimatake H, Court D, Schmeissner U, Brady C, Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Duckworth ML, Gait MJ, Goelet P, Hong GF, Singh M, Titmas RC. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. [PMC free article] [PubMed]
  • Mackie GA. Nucleotide sequence of the gene for ribosomal protein S20 and its flanking regions. J Biol Chem. 1981 Aug 10;256(15):8177–8182. [PubMed]
  • Poulis MI, Shaw DC, Campbell HD, Young IG. In vitro synthesis of the respiratory NADH dehydrogenase of Escherichia coli. Role of UUG as initiation codon. Biochemistry. 1981 Jul 7;20(14):4178–4185. [PubMed]
  • De Robertis EM, Jr, Judewicz RD, Torres HN. On the control mechanism of bacterial growth by cyclic adenosine 3',5'-monophosphate. Biochem Biophys Res Commun. 1973 Dec 10;55(3):758–764. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Gene
    Gene links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...