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Studies on the involvement of the UGA readthrough process in the mechanism of attenuation of the tryptophan operon of Escherichia coli

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Summary

We asked if UGA suppression by charged tRNATrp, a process called UGA readthrough, is involved in the mechanism of attenuation of the tryptophan (trp) operon in Escherichia coli. For this purpose we used two mutations: strA (LD1) which causes restriction of UGA readthrough, and revA which partially overcomes the restriction of UGA readthrough caused by strA (LD1) (Engelberg-Kulka et al. 1982). trp attenuation was monitored by the regulation of the synthesis of the trp operon enzyme anthranilate synthetase (ASase) in trpR- strains. We showed that the strA (LD1) mutation causes a significant increase in the level of synthesis of ASase in the presence of an excess of tryptophan, while the revA mutation reverses this effect, indicating that transcription termination at the trp attenuator site is relieved by restriction of UGA readthrough. Based on our results and the sequence data of the trp leader RNA of E. coli (Oxender et al. 1979), we offer a model for the involvement of the UGA readthrough process in trp attenuation. We suggest that the UGA readthrough process permits trp attenuation to respond to slight changes in the cellular concentration of charged tRNATrp.

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References

  • Adler HI, Fisher WD, Cohen A, Hardigree AA (1967) Miniatur Escherichia coli cells deficient in DNA. Proc Natl Acad Sci USA 57:321–326

    Google Scholar 

  • Barbour SD, Nagaishi A, Templin A, Clark AJ (1970) Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants caused by indirect suppression of Rec- mutations. Proc Natl Acad Sci USA 67:128–135

    Google Scholar 

  • Bennett GN, Schweingruber ME, Brown KD, Squires C, Yanofsky C (1976) Nucleotide sequence of region preceding trp mRNA initiation site and its role in promoter and operator function. Proc Natl Acad Sci USA 73:2351–2355

    Google Scholar 

  • Bennett GN, Schweingruber ME, Brown KD, Squires C, Yanofsky C (1978) Nucleotide sequence of the promoter-operator region of the tryptophan operon of Escherichia coli. J Mol Biol 121:113–137

    Google Scholar 

  • Bennett GN, Yanofsky C (1978) Sequence analysis of operator constitutive mutants of the tryptophan operon of Escherichia coli. J Mol Biol 121:179–192

    Google Scholar 

  • Bertrand K, Korn L, Lee F, Platt T, Squires CL, Squires C, Yanofsky C (1975) New features of the regulation of the tryptophan operon. Science 189:22–26

    Google Scholar 

  • Bertrand K, Squires C, Yanofsky C (1976) Transcription termination in vivo in the leader region of the tryptophan operon of Escherichia coli. J Mol Biol 103:319–337

    Google Scholar 

  • Davis BD, Mingioli EJ (1950) Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol 60:17–28

    Google Scholar 

  • DiNocera PP, Blasi F, DiLaru R, Frunzio R, Bruni CB (1978) Nucleotide sequence of the attenuator region of the histidine operon of Escherichia coli K-12. Proc Natl Acad Sci USA 75:4276–4280

    Google Scholar 

  • Engelberg-Kulka H, Amiel A, Dekel L, Raveh B, Schoulaker-Schwarz R (1982) Genetic analysis of a streptomycin-resistant Escherichia coli mutant temperature-sensitive for nonsense suppression. Mol Gen Genet 188:149–155

    Google Scholar 

  • Engelberg-Kulka H, Dekel L, Israeli-Reches M (1977) Streptomycin-resistant Escherichia coli mutant temperature-sensitive for the production of Qβ-infective particles. J Virol 21:1–6

    Google Scholar 

  • Engelberg-Kulka, Dekel L, Israeli-Reches M, Belfort M (1979) The requirement of nonsense suppression for the development of several phages. Mol Gen Genet 170:155–159

    Google Scholar 

  • Engelberg-Kulka H (1981) UGA suppression by normal tRNATrp in Escherichia coli. Codon context effect. Nucl Acids Res 9:983–991

    Google Scholar 

  • Engelberg-Kulka H, Dekel L, Israeli-Reches M (1981) Regulation of the Escherichia coli tryptophan operon by readthrough of UGA termination codons. Biochem Biophys Res Commun 98:1008–1015

    Google Scholar 

  • Gardner JF (1979) Regulation of the threonine operon. Tandem threonine and isoleucine codons in the control region and translational control of transcription termination. Proc Natl Acad Sci USA 76:1700–1710

    Google Scholar 

  • Hirsh D (1971) Tryptophan transfer RNA as the UGA suppressor. J Mol Biol 58:439–458

    Google Scholar 

  • Ito J, Crawford IP (1965) Regulation of the enzymes of the tryptophan pathway in Escherichia coli. Genetics 52:1303–1316

    Google Scholar 

  • Jackson EN, Yanofsky C (1973) The region between the operator and the first structural gene of the tryptophan operon of Escherichia coli may have a regulatory function. J Mol Biol 76:89–101

    Google Scholar 

  • Lawther RP, Hatfield GW (1980) Multivalent translational control of transcription termination at attenuator of ilv GEDA operon of Escherichia coli K-12. Proc Natl Acad Sci USA 77:1862–1866

    Google Scholar 

  • Lee F, Yanofsky C (1977) Transcription termination of the trp operon attenuators of Escherichia coli and Salmonella typhimurium: RNA secondary structure and regulation of termination. Proc Natl Acad Sci USA 74:4365–4369

    Google Scholar 

  • McGeoch D, McGeoch J, Morse D (1973) Synthesis of tryptophan operon RNA in cell-free system. Nature (London) New Biol 245:137–140

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, pp 230–234

  • Moore CH, Farron F, Bohnert D, Weissmann C (1971) Possible origin of a minor virus specific protein (A1) in Qβ particles. Nature (London) New Biol 234:204–206

    Google Scholar 

  • Morse DE, Morse ANC (1976) Transcription termination in vivo in the leader region of the tryptophan operon of Escherichia coli. J Mol Biol 103:319–337

    Google Scholar 

  • Morse DE, Yanofsky C (1968) Amber mutants of the trR regulatory gene. J Mol Biol 44:185–193

    Google Scholar 

  • Nargang FE, Subrahmanyam CS, Umbarger HE (1980) Nucleotide sequence of ilvGEDA operon attenuator region of Escherichia coli. Proc Natl Acad Sci USA 77:1823–1827

    Google Scholar 

  • Oxender DL, Zurawski G, Yanofsky C (1979) Attenuation in the Escherichia coli tryptophan operon. Role of RNA secondary structure involving the tryptophan codon region. Proc Natl Acad Sci USA 76:5524–5528

    Google Scholar 

  • Rose JK, Yanofsky C (1974) Interaction of the operator of the tryptophan operon with repressor. Proc Natl Acad Sci USA 71:3134–3138

    Google Scholar 

  • Squires CL, Lee F, Yanofsky C (1975) Interaction of the trp repressor and RNA polymerase with the trp operon. J Mol Biol 92:93–111

    Google Scholar 

  • Squies CL, Rose JK, Yanofsky C, Yang HL, Zubay G (1973) Regulation of in vitro transcription of the tryptophan operon by purified RNA polymerase in the presence of partially purified repressor and tryptophan. Nature (London) New Biol 245:133–137

    Google Scholar 

  • Weiner AM, Weber K (1971) Natural readthrough at the UGA termination signal of Qβ coat protein cistron. Nature (London) New Biol 234:206–209

    Google Scholar 

  • Yanofsky C, Soll L (1977) Mutations affecting tRNATrp and its charging and their effect on regulation of transcription termination at the attenuator of the tryptophan operon. J Mol Biol 113:663–677

    Google Scholar 

  • Yanofsky C (1981) Attenuation in the control of expression of bacterial operons. Nature (London) 289:751–758

    Google Scholar 

  • Zeevi M, Daniel V, Engelberg-Kulka H (1979) A streptomycinresistant Escherichia coli mutant with ribosomes temperaturesensitive in suppression of a nonsense codon. Mol Gen Genet 170:149–153

    Google Scholar 

  • Zurawski G, Brown K, Killingly D, Yanofsky C (1978) Nucleotide sequence of the leader region of the phenylalanine operon of Escherichia coli. Proc Natl Acad Sci USA 75:4271–4275

    Google Scholar 

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Authors and Affiliations

  1. Department of Molecular Biology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel

    Hanna Engelberg-Kulka, Aliza Amiel, Chaya Miller & Rachel Schoulaker-Schwarz

Authors
  1. Hanna Engelberg-Kulka
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  2. Aliza Amiel
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  3. Chaya Miller
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  4. Rachel Schoulaker-Schwarz
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Communicated by K. Isono

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Engelberg-Kulka, H., Amiel, A., Miller, C. et al. Studies on the involvement of the UGA readthrough process in the mechanism of attenuation of the tryptophan operon of Escherichia coli . Molec Gen Genet 188, 156–160 (1982). https://doi.org/10.1007/BF00333011

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