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Constitutive expression in Escherichia coli of the Neurospora crassa structural gene encoding the inducible enzyme catabolic dehydroquinase

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Summary

In Neurospora crassa the qa-2 gene, which encodes catabolic dehydroquinase, is under positive control exerted by the inducer quinic acid and an activator protein encoded in the closely linked qa-1 gene. In order to determine if this regulatory mechanism is maintained when the qa-2 gene is cloned on a recombinant plasmid and expressed in Escherichia coli, molecular cloning experiments have been performed using DNA isolated from a qa-1 +(inducible), a qa-1C (constitutive) and two qa-1 (non-inducible) strains of N. crassa. The results demonstrate that the level of expression of the qa-2 gene in E. coli is completely independent of the mutational state of the qa-1 gene. Moreover, the level of expression of the cloned qa-2 gene was unaffected by either an intracellularly produced inducer of catabolic dehydroquinase or by the general procaryotic positive effector, the CAP factor. The weight of evidence thus supports the conclusion that transcription of the N. crassa qa-2 gene in E. coli does not require the qa-1 activator protein and thus is not controlled by the same mechanism which functions in N. crassa.

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References

  • Alton, N.K., Hautala, J.A., Giles, N.H., Kushner, S.R., Vapnek, D.: Transcription and translation in E. coli of hybrid plasmids containing the catabolic dehydroquinase gene from Neurospora crassa. Gene 4, 241–259 (1978)

    Google Scholar 

  • Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Bacteriol. Rev. 40, 116–167 (1976)

    Google Scholar 

  • Carbon, J., Ratzkin, B., Clarke, L., Richardson, D.: The expression of cloned DNA in prokaryotes. Brookhaven Symp. Biol. 29, 277–297 (1977)

    Google Scholar 

  • Case, M.E., Giles, N.H.: Genetic evidence on the organization and action of the qa-1 gene product: a protein regulating the induction of three enzymes in quinate catabolism in Neurospora crassa. Proc. Natl. Acad. Sci. U.S.A. 72, 553–557 (1975)

    Google Scholar 

  • Case, M.E., Giles, N.H., Doy, C.H.: Genetical and biochemical evidence for further interrelationships between the polyaromatic synthetic and the quinate-shikimate catabolic pathways in Neurospora crassa. Genetics 71, 337–348 (1972)

    Google Scholar 

  • Chaleff, R.S.: The inducible quinate-shikimate catabolic pathway in Neurospora crassa: genetic organization. J. Gen. Microbiol. 81, 337–355 (1974)

    Google Scholar 

  • Dickson, R.C., Markin, J.S.: Molecular cloning and expression in E. coli of a yeast gene coding for β-galactosidase. Cell 15, 123–130 (1974)

    Google Scholar 

  • Dickson, R. C., Markin, J.S.: Molecular cloning and expression in E. coli of a yeast gene coding for β-galactosidase. Cell 15, 123–130 (1978)

    Google Scholar 

  • Englesberg, E., Wilcox, G.: Regulation: positive control. In: Ann. Rev. Genetics (Roman, Campbell and Sandler, eds.), pp. 219–242 (1974)

  • Hautala, J.A., Jacobson, J.W., Case, M.E., Giles, N.H.: Purification and characterization of catabolic dehydroquinase, an enzyme in the inducible quinic acid catabolic pathway of Neurospora crassa. J. Biol. Chem. 250, 6008–6014 (1975)

    Google Scholar 

  • Jacobson, J.W., Hautala, J.A., Lucas, M.C., Reinert, W.R., Strøman, P., Barea, J.L., Patel, V.B., Case, M.E., Giles, N.H.: Regulation of quinate catabolism in Neurospora: the qa gene cluster. In: Molecular approaches to eucaryotic genetic systems (ICN-UCLA Symposia on Molecular and Cellular Biology, Vol. VIII), pp. 269–283. New York: Academic Press 1977

    Google Scholar 

  • Kleckner, N., Roth, J., Botstein, D.: Genetic engineering in vivo using translocatable drug-resistance elements. J. Mol. Biol. 116, 125–159 (1977)

    Google Scholar 

  • Kushner, S.R.: An improved method for transformation of Escherichia coli with ColEl derived plasmids. In: Genetic engineering (H.W. Boyer and S. Nicosia, eds.), pp. 23–47. Amsterdam, The Netherlands: Elsevier/North Holland Biomedical Press 1978

    Google Scholar 

  • Kushner, S.R., Hautala, J.A., Jacobson, J.W., Giles, N.H., Vapnek, D.: Expression of the structural gene for catabolic dehydroquinase of Neurospora crassa in Escherichia coli K12. Brookhaven Symp. Biol. 29, 297–308 (1977)

    Google Scholar 

  • Ratzkin, Carbon, J.: Functional expression of cloned yeast DNA in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 74, 487–491 (1977)

    Google Scholar 

  • Rines, H.W., Case, M.E., Giles, N.H.: Mutants in the arom gene cluster of Neurospora crassa specific for biosynthetic dehydroquinase. Genetics 61, 789–800 (1968)

    Google Scholar 

  • Struhl, K., Cameron, J.R., Davis, R.W.: Functional expression of eukaryotic DNA in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 73, 1471–1475 (1976)

    Google Scholar 

  • Valone, J.A., Case, M.E., Giles, N.H.: Constitutive mutants in a regulatory gene exerting positive control of quinic acid catabolism in Neurospora crassa. Proc. Natl. Acad. Sci. U.S.A. 68, 1555–1559 (1971)

    Google Scholar 

  • Vapnek, D., Hautala, J.A., Jacobson, J.W., Giles, N.H., Kushner, S.R.: Expression in Escherichia coli K-12 of the structural gene for catabolic dehydroquinase of Neurospora crassa. Proc. Natl. Acad. Sci. U.S.A. 74, 3508–3512 (1977)

    Google Scholar 

  • White, B.J., Hochhauser, S.J., Cintron, N.M., Weiss, B.: Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12. J. Bacteriol. 126, 1082–1088 (1976)

    Google Scholar 

  • Zeig, J., Maples, V.F., Kushner, S.R.: Recombination levels of Escherichia coli K-12 mutants deficient in various replication, recombination, or repair genes. J. Bacteriol. 134, 958–966 (1978)

    Google Scholar 

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Communicated by W. Gehring

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Hautala, J.A., Schweizer, M., Giles, N.H. et al. Constitutive expression in Escherichia coli of the Neurospora crassa structural gene encoding the inducible enzyme catabolic dehydroquinase. Molec. Gen. Genet. 172, 93–98 (1979). https://doi.org/10.1007/BF00276219

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