Abstract
The coordinate expression of four sequential enzymes in the de novo pyrimidine pathway may result from the interaction of the various polypeptides of the pathway in Serratia marcescens rather than represent some unit of transcriptional regulation. These interactions were defined by examining the polypeptide association observed in extracts of parental and mutant strains in a series of pleiotropic pyrimidine auxotrophs. Extracts of pyrE auxotrophs [possessing dihydroorotate (DHOase) activity but no orotidine-5′-monophosphate pyrophosphorylase (OMPppase) activity] stimulate OMPppase activity in extracts of pyrC auxotrophs (possessing reduced OMPppase activity but no DHOase activity). Separation by molecular weight on Sephadex G200 has suggested an aggregation between the final two enzymes, OMPppase and OMPdecarboxylase (OMPdecase), and the earlier enzyme, DHOase. The reduction of OMPppase activity in pyrC auxotrophs (encoding either a defective polypeptide or reduced levels) is explained by the lack of adequate levels of DHOase for aggregate formation. Such polypeptide interactions appear to mimic the coordinate formation of polypeptides which are controlled as a unit of regulation. The measurable levels of enzymatic activity vary in a quantitatively identical manner, but the variation does not result directly from the regulation of polypeptide formation.
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References
Adelberg, E. A., Mandel, M., and Chen, G. C. C. (1965). Optimal conditions for mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem. Biophys. Res. Commun. 18788.
Ames, B. N., and Garry, B. J. (1959). Coordinate repression of the synthesis of four histidine biosynthetic enzymes by histidine. Proc. Natl. Acad. Sci. 451453.
Ames, B. N., Garry, B., and Herzenberg, L. A. (1960). The genetic control of the enzymes of histidine biosynthesis in Salmonella typhimurium. J. Gen. Microbiol. 22369.
Bachmann, B. J., Low, K. B., and Taylor, A. L. (1976). Recalibrated linkage map of Escherichia coli K-12. Bacteriol. Rev. 40116.
Beckwith, J. R., Pardee, A. B., Austrian, R., and Jacob, F. (1962). Coordination of the synthesis of the enzymes in the pyrimidine pathway. J. Mol. Biol. 5618.
Blair, D. G. R., and Potter, V. R. (1961). Inhibition of orotidylic acid decarboxylase by uridine 5′-phosphate. J. Biol. Chem. 2362503.
Calvo, J. M., and Fink, G. R. (1971). Regulation of biosynthetic pathways in bacteria and fungi. Ann. Rev. Biochem. 40943.
Condon, S., Condon, J. K., and O'Donovan, G. A. (1976). Regulation of arginine and pyrimidine biosynthesis in Pseudomonas putida. J. Gen. Microbiol. 92375.
Cunin, R., Elsebiers, D., Sand, G., Freundlick, G., and Glansdorff, N. (1969). On the functional organization of the arg ECBH cluster of genes in Escherichia coli K-12. Mol. Gen. Genet. 10632.
DeMoss, J. A. (1965). Biochemical diversity in the tryptophan pathway. Biochem. Biophys. Res. Commun. 18850.
Dennis, P. P., and Herman, R. K. (1970). Pyrimidine pools and macromolecular composition of pyrimidine-limited Escherichia coli. J. Bacteriol. 102118.
Frieden, C. (1971). Protein-protein interaction and enzymatic activity. Ann. Rev. Biochem. 40653.
Gerhart, J. C., and Pardee, A. B. (1962). The enzymology of control by feedback inhibition. J. Biol. Chem. 237891.
Ginsburg, A., and Stadtman, E. R. (1970). Multienzyme systems. Ann. Rev. Biochem. 39429.
Hayward, W. S., and Belser, W. L. (1965). Regulation of pyrimidine biosynthesis in Serratia marcescens. Proc. Natl. Acad. Sci. 531483.
Hutchinson, M. A., and Belser, W. L. (1969). Enzymes of tryptophan biosynthesis in Serratia marcescens. J. Bacteriol. 98109.
Hutson, J. Y., and Downing, M. (1968). Pyrimidine biosynthesis in Lactobacillus leichmanii. J. Bacteriol. 961249.
Issac, J. H., and Holloway, B. W. (1968). Control of pyrimidine biosynthesis in Pseudomonas aeruginosa. J. Bacteriol. 961732.
Jones, M. E. (1972). Regulation of uridylic acid biosynthesis in eukaryotic cells. Curr. Top. Cell. Regul. 6227.
Karibian, D., and Couchoud, P. (1974). Dihydro-orotate oxidase of Escherichia coli K12: Purification, properties, and relation to the cytoplasmic membrane Biochim. Biophys. Acta 364218.
Kelleher, R. J., and Gooder, H. (1973). Genetics and biochemistry of pyrimidine biosynthesis in Bacillus subtilis: Linkage between mutations resulting in a requirement for uracil. J. Bacteriol. 116577.
Kelln, R. A., Kinahan, J. J., Foltermann, K. F., and O'Donovan, G. A. (1975). Pyrimidine biosynthetic enzymes in Salmonella typhimurium, repressed specifically by growth in the presence of cytidine. J. Bacteriol. 124764.
Labrum, E. L., and Bunting, M. I. (1953). Spontaneous and induced color variation of the HY strain of Serratia marcescens, J. Bacteriol. 65394.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurements with the Folin phenol reagent. J. Biol. Chem. 193265.
Maas, W. K. (1961). Studies on repression of arginine biosynthesis in Escherichia coli. Cold Spring Harbor Symp. Quant. Biol. 26183.
Maas, W. K., and McFall, E. (1964). Genetic aspects of metabolic control. Ann. Rev. Microbiol. 1895.
O'Donovan, G. A., and Neuhard, J. (1970). Pyrimidine metabolism in microorganisms. Bacteriol. Rev. 34278.
Potvin, B. W., Kelleher, R. J., Jr., and Gooder, H. (1975). Pyrimidine biosynthetic pathway of Bacillus subtilis. J. Bacteriol. 123604.
Sanderson, K. E. (1972). Linkage map of Salmonella typhimurium: Edition IV. Bacteriol. Rev. 36558.
Schwartz, M., and Neuhard, J. (1975). Control of expression of the pyr genes in Salmonella typhimurium: Effects of variations in uridine and cytidine nucleotide pools. J. Bacteriol. 121814.
Shoaf, W. T., and Jones, M. E. (1973). Uridylic acid synthesis in Ehrlich ascites carcinoma: Properties, subcellular distribution, and nature of enzyme complexes of the six biosynthetic enzymes. Biochemistry 124039.
Welch, G. R., and Gaertner, F. H. (1976). Coordinate activation of a multienzyme complex by the first substrate. Arch. Biochem. Biophys. 172476.
Wild, J. R., and Belser, W. L. (1977). Pyrimidine biosynthesis in Serratia marcescens: Polypeptide interactions of three nonsequential enzymes. Biochem. Genet. 15173.
Wild, J. R., Belser, W. L., and O'Donovan, G. A. (1976). The aspartate transcarbamylase of Serratia marcescens HY. J. Bacteriol. 128766.
Williams, J. C., and O'Donovan, G. A. (1973). Repression of enzyme synthesis of the pyrimidine pathway in Salmonella typhimurium. J. Bacteriol. 1151071.
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This work was supported by grants from the National Science Foundation (NSF GB 5811) and the Office of Naval Research (Nonr 4413). One of us (J.W.) was a National Science Foundation Graduate Fellow.
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Wild, J.R., Belser, W.L. Pyrimidine biosynthesis in Serratia marcescens: A possible role for nonsequential enzyme interactions in mimicking coordinate gene expression. Biochem Genet 15, 157–172 (1977). https://doi.org/10.1007/BF00484559
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DOI: https://doi.org/10.1007/BF00484559