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.
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)
Bergmeyer, H-U.: Methods of enzymatic analysis. 2nd English Edit. Weinheim/Bergstr. and, London and New York: Verlag Chemie and Academic Press 1974
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
Bratton, A.C., Marshall, E.K.: A new coupling component for sulphanilamide determination. J. biol. Chem.128, 537–550 (1939)
Bücher, T.: Über ein phosphatübertragendes Gärungsferment. Biochim. biophys. Acta (Amst.),1, 292–394 (1974)
Costello, W.P., Bevan, E.A.: Complementation betweenade5,7 alleles in yeast. Genetics50, 1219–1230 (1964)
Dorfman, B.: Allelic complementation at theade5,7 locus in yeast. Genetics50, 1231–1243 (1964)
Duntze, W., Manney, T.R.: Two mechanisms for allelic complementation among tryptophan synthetase mutants ofSaccharomyces cerevisiae. J. Bact.96, 2085–2093 (1968)
Fincham, J.R.S.: Genetic complementation. New York: Benjamin 1966
Fisher, C.R.: Enzymology of the pigmented adenine-requiring mutants ofSaccharomyces andSchizosaccharomyces. Biochem. biophys. Res. Commun.34, 306–310 (1969)
Flaks, J.G., Lukens, L.N.: The enzymes of purine nucleotide synthesisde novo. Methods in Enzymology6, 52–95 (1963)
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)
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
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)
Henderson, J.F.: Regulation of purine biosynthesis. Washington: American Chemical Society 1972
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)
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)
Lukens, I.N., Flaks, J.G.: Intermediates in purine nucleotide synthesis. Methods in Enzymology6, 671–702 (1963)
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)
Megmet, R., Giles, N.H.: Allelic complementation at the adenylosuccinase locus inSchizosaccharomyces pombe. Genetics50, 967–971 (1964)
Nicholas, D.J., Nason, A.: Determination of nitrate and nitrite. Methods in Enzymology3, 981–984 (1957)
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)
Sanderson, K.E.: Current linkage map ofSalmonella typhimurium. Bact. Rev.34, 176–193 (1970)
Taylor, A.L.: Current linkage map ofEscherichia coli. Bact. Rev.34, 155–175 (1970)
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
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)
Wanner, H.: Die Akkumulation von Hypoxanthin durch adeninabhängige Mutanten vonS. pombe. Arch. Klaus-Stift. Zürich30, 516–520 (1955)
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)
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)
Wyss, J.: Enzymatischer analyse adenin-auxotropher Mutanten vonSchizosaccharomyces pombe. Untersuchung des Enzyms Glycinamid ribotid-transformylase. Lizentiatsarbeit, Universität Bern, Switzerland. (1971)
Yourno, J.D.: Gene fusion. In: Evolution of genetic systems, H.H. Smith (ed.), pp. 95–120. New York, Gordon and Breach 1972
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)
Author information
Authors and Affiliations
Additional information
Communicated by F. Kaudewitz
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00582878