Skip to main content
Log in

The product of theade1 gene inSchizosaccharomyces pombe: A bifunctional enzyme catalysing two distinct steps in purine biosynthesis

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Summary

The assignment of the knownade genes to steps in purine biosynthesis inSchizosaccharomyces pombe has been completed with the demonstration that anade3 mutants lacks FGAR amidotransferase,ade1A mutants lack GAR synthetase andade1B mutants lack AIR synthetase. A comparison of enzyme activity with map position forade1 mutants shows that (1) complementingade1A mutants lack GAR synthetase but possess wild type amounts of AIR synthetase, (2) complementingade1B mutants lack AIR synthetase but posses variable amounts of GAR synthetase, (3) non-complementing mutants lack both activities. In wild type strains the two activities fractionate together throughout a hundred-fold purification. Hence theade1 gene appears to code for a bifunctional enzyme catalysing two distinct steps in purine biosynthesis. The two activities are catalysed by two different regions of the polypeptide chain which can be altered independendently by mutation. Gel filtration studies on partially purified enzymes from wild type and various complementing mutant strains, indicate that the bifunctional enzyme is a multimer consisting of between four and six sub-units of 40,000 daltons each. GAR synthetase activity is associated with both the monomeric and multimeric forms but AIR synthetase is only associated with the multimer. A comparison of enzyme levels between diploids and their original complementing haploid strains suggests that complementation is due to hybrid enzyme formation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Andrews, P.: The gel filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J.96, 595–606 (1965)

    Google Scholar 

  • Bergmeyer, H-U.: Methods of enzymatic analysis. 2nd English Edit. Weinheim/Bergstr. and, London and New York: Verlag Chemie and Academic Press 1974

    Google Scholar 

  • Bonner, D.M., DeMoss, J.A., Mills, S.E.: The evolution of an enzyme. In: Evolving genes and proteins, Bryson, V. and Vogel, H.J. (eds.). New York and London: Academic Press 1965

    Google Scholar 

  • Bratton, A.C., Marshall, E.K.: A new coupling component for sulphanilamide determination. J. biol. Chem.128, 537–550 (1939)

    Google Scholar 

  • Bücher, T.: Über ein phosphatübertragendes Gärungsferment. Biochim. biophys. Acta (Amst.),1, 292–394 (1974)

    Google Scholar 

  • Costello, W.P., Bevan, E.A.: Complementation betweenade5,7 alleles in yeast. Genetics50, 1219–1230 (1964)

    Google Scholar 

  • Dorfman, B.: Allelic complementation at theade5,7 locus in yeast. Genetics50, 1231–1243 (1964)

    Google Scholar 

  • Duntze, W., Manney, T.R.: Two mechanisms for allelic complementation among tryptophan synthetase mutants ofSaccharomyces cerevisiae. J. Bact.96, 2085–2093 (1968)

    Google Scholar 

  • Fincham, J.R.S.: Genetic complementation. New York: Benjamin 1966

    Google Scholar 

  • Fisher, C.R.: Enzymology of the pigmented adenine-requiring mutants ofSaccharomyces andSchizosaccharomyces. Biochem. biophys. Res. Commun.34, 306–310 (1969)

    Google Scholar 

  • Flaks, J.G., Lukens, L.N.: The enzymes of purine nucleotide synthesisde novo. Methods in Enzymology6, 52–95 (1963)

    Google Scholar 

  • Friis, J., Flury, F., Leupold, U.: Characterisation of spontaneous mutants of mitotic and meiotic origin in theade1 locus ofSchizosaccharomyces pombe. Mutation Res.11, 373–390 (1971)

    Google Scholar 

  • Gutz, H., Heslot, H., Leupold, U., Loprieno, N.:Schizosaccharomyces pombe. In: Handbook of genetics, R.C. King (ed.), Vol. 1, pp. 395–446. New York and London: Plenum Press 1974

    Google Scholar 

  • Gross, T.S., Woods, R.A.: Identification of mutants defective in the first and second steps ofde novo purine synthesis inSaccharomyces cerevisiae. Biochim. biophys. Acta (Amst.)247 13–21 (1971)

    Google Scholar 

  • Henderson, J.F.: Regulation of purine biosynthesis. Washington: American Chemical Society 1972

    Google Scholar 

  • Heslot, H., Nagy, M., Whitehead, E.: Rechérches génétiques et biochimiques sur la première enzyme de la biosynthèse des purines chez leSchizosaccharomyces pombe. C.R. Acad. Sci. (Paris), Ser. D Sci. Nat.263, 57–58 (1966)

    Google Scholar 

  • Lowry, O.H., Roseborough, N.J., Farr A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951)

    Google Scholar 

  • Lukens, I.N., Flaks, J.G.: Intermediates in purine nucleotide synthesis. Methods in Enzymology6, 671–702 (1963)

    Google Scholar 

  • Manney, T.R., Duntze, W., Janosko, N., Salazar, J.: Genetical and biochemical studies of partially active tryptophan synthetase mutants ofSaccharomyces cerevisiae. J. Bact.99, 590–596 (1969)

    Google Scholar 

  • Megmet, R., Giles, N.H.: Allelic complementation at the adenylosuccinase locus inSchizosaccharomyces pombe. Genetics50, 967–971 (1964)

    Google Scholar 

  • Nicholas, D.J., Nason, A.: Determination of nitrate and nitrite. Methods in Enzymology3, 981–984 (1957)

    Google Scholar 

  • Nierlich, D.P., Magasanik, B.: Phosphoribosylglycineamide synthetase ofAerobacter aerogenes. Purification and properties and non-enzymatic formation of its substrate 5′-phosphoribosylamine. J. biol. Chem.240, 366–374 (1965)

    Google Scholar 

  • Sanderson, K.E.: Current linkage map ofSalmonella typhimurium. Bact. Rev.34, 176–193 (1970)

    Google Scholar 

  • Taylor, A.L.: Current linkage map ofEscherichia coli. Bact. Rev.34, 155–175 (1970)

    Google Scholar 

  • Thomas, C.A., Abelson, J.: The isolation and characterisation of DNA from bacterophage. In: Procedures in nucleic acid research, Cantoni, G.L. and Davies D.R. (eds.), pp 553–561. New York: Harper and Row 1966

    Google Scholar 

  • Tritz, G.J., Matney, T.S., Chandler, J.L.R., Gholson, R.K.: Identification of thepurI locus inEscherichia coli K12. J. Bact.102, 881–883 (1970)

    Google Scholar 

  • Wanner, H.: Die Akkumulation von Hypoxanthin durch adeninabhängige Mutanten vonS. pombe. Arch. Klaus-Stift. Zürich30, 516–520 (1955)

    Google Scholar 

  • Westby, C.A., Gots, J.S.: Genetic blocks and unique features of 5-phosphoribosyl-N-formylglycineamide biosynthesis inSalmonelle typhimurium. J. biol. Chem.244, 2095–2102 (1969)

    Google Scholar 

  • Whitehead, E., Nagy, M., Heslot, H.: Interactions entre la biosynthèse des purines et celles de l'histidine chez leSchizosaccharomyces pombe. C.R. Acad. Sci. (Paris), Ser. D Sci. Nat.263, 819–821 (1966)

    Google Scholar 

  • Wyss, J.: Enzymatischer analyse adenin-auxotropher Mutanten vonSchizosaccharomyces pombe. Untersuchung des Enzyms Glycinamid ribotid-transformylase. Lizentiatsarbeit, Universität Bern, Switzerland. (1971)

    Google Scholar 

  • Yourno, J.D.: Gene fusion. In: Evolution of genetic systems, H.H. Smith (ed.), pp. 95–120. New York, Gordon and Breach 1972

    Google Scholar 

  • Yourno, J.D., Kohno, T., Roth, J.R.: Enzyme evolution: generation of a bifunctional enzyme by fusion of adjacent genes. Nature (Lond.)228, 820–824 (1970)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by F. Kaudewitz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fluri, R., Coddington, A. & Flury, U. The product of theade1 gene inSchizosaccharomyces pombe: A bifunctional enzyme catalysing two distinct steps in purine biosynthesis. Molec. Gen. Genet. 147, 271–282 (1976). https://doi.org/10.1007/BF00582878

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00582878

Keywords

Navigation