Abstract
We have characterized biochemical effects of Idh GB1in Drosophila melanogaster. This is a “null”-activity allele for NADP+-dependent isocitrate dehydrogenase (NADP-IDH) isolated from a natural population. The homozygous mutant strain has 5% of the NADP-IDH specific activity found in controls and less than 24% of the immunologically cross-reacting material (CRM). This mutation maps to 27.2 on the third chromosome, to the right of h. The biochemical phenotype of this mutant strain includes a coordinate reduction in malic enzyme (ME) specific activity and CRM and an increase in specific activity for the pentose-phosphate shunt enzymes, 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase. The K m values for purified NADP-IDH are not different from those found for the purified control enzyme for NADP+ or isocitrate. It is suggested that this allele may represent a cis-acting control mutation for one of at least two loci involved in the production of NADP-IDH in D. melanogaster.
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References
Bentley, M. M. (1983). Linkage data. Dros. Inform. Serv. 59 (in press).
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principles of protein-dye binding. Anal. Biochem. 72248.
Fox, D. J. (1971). The soluble citric acid cycle enzymes of Drosophila melanogaster. I. Genetics and ontogeny of NADP-linked isocitrate dehydrogenase. Biochem. Genet. 569.
Geer, B. W., Krochko, D., and Williamson, J. H. (1979a). Ontogeny, cell distribution, and the physiological role of NADP-malic enzyme in Drosophila melanogaster. Biochem. Genet. 17867.
Geer, B. W., Lindel, D. L., and Lindel, D. M. (1979b). Relationship of the oxidative pentose shunt pathway to lipid synthesis in Drosophila melanogaster. Biochem. Genet. 17881.
Geer, B. W., Krochko, D., Oliver, M. J., Walker, V. K., and Williamson, J. H. (1980). A comparative study of the NADP-malic enzymes from Drosophila and chick liver. Comp. Biochem. Physiol. 65B25.
Geer, B. W., Williamson, J. H., Cavener, D. R., and Cochrane, B. J. (1981). Dictary modulation of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in Drosophila. In Bhaskaran, G., Friedman, S., and Rodriguez, J. G. (eds.), Current Topics in Insect Endocrinology and Nutrition Plenum, New York, pp. 253–281.
Glock, G. E., and McLean, P. (1953). Further studies on the properties and assay of glucose-6-phosphate dehydrogenase of rat liver. Biochem. J. 55400.
Horie, Y. (1967). Dehydrogenase in carbohydrate metabolism in larvae of the silkworm, Bombyx mori L. J. Insect Physiol. 131163.
Langley, C. H., Voelker, R. A., Leigh Brown, A. J., Ohnishi, S., Dickson, B., and Montgomery, E. (1981). Null allele frequencies at allozyme loci in natural populations of Drosophila melanogaster. Genetics 99151.
Laurell, C. B. (1966). Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal. Biochem. 1545.
Lewis, E. B. (1960). A new standard food medium. Dros. Inform. Serv. 34117.
Lindsley, D. L., and Grell, E. H. (1968). Genetic Variations of Drosophila melanogaster, Carnegie Inst. Wash. Publ. No. 627.
Ochoa, S. (1955). Malic enzyme. In Colourik, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. 1 Academic Press, New York, pp. 739–753.
Ohnishi, S., and Voelker, R. A. (1979). Comparative studies of allozyme loci in Drosophila simulans and D. melanogaster. II. Gene arrangement on the third chromosome. Jap. J. Genet. 54203.
Ohnishi, S., and Voelker, R. A. (1982). New data on the genetic mapping of isocitrate dehydrogenase in D. melanogaster. Dros. Inform. Serv. 58121.
Rawls, J. M., Jr., and Lucchesi, J. C. (1974). Regulation of enzyme activities in Drosophila I. The detection of regulatory loci by gene dosage responses. Genet. Res. Cambr. 2459.
Rohlf, F. J., and Sokal, R. R. (1969). Statistical Tables W. H. Freeman, San Francisco.
Sokal, R. R., and Rohlf, F. J. (1969). Biometry W. H. Freeman, San Francisco.
Stewart, B. R., and Merriam, J. R. (1974). Segmental aneuploidy and enzyme activity as a method for cytogenetic localization in Drosophila melanogaster. Genetics 76301.
Ward, C. L., and Alexander M. L. (1957). Cytological analysis of X-ray-induced mutations at eight specific loci in the third chromosome of Drosophila melanogaster. Genetics 4242.
Williamson, J. H. (1982). Men NCI: A putative regulatory mutant of NADP-Malic enzyme in Drosophila melanogaster. Can. J. Genet. Cytol. 24409.
Williamson, J. H., and Bentley, M. M. (1983). Dosage compensation in Drosophila: NADP-enzyme activities and cross-reacting material. Genetics 103649.
Williamson, J. H., Krochko, D., and Bentley, M. M. (1980). Properties of Drosophila NADP+-isocitrate dehydrogenase purified on Procion Brilliant Blue-Sepharose-4B. Comp. Biochem. Physiol. 65B339.
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Research supported by an Alberta Heritage Foundation for Medical Research Establishment Grant to MMB and a Natural Sciences and Engineering Research Council Operating Grant to JHW.
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Bentley, M.M., Meidinger, R.G. & Williamson, J.H. Characterization of a low-activity allele of NADP+-dependent isocitrate dehydrogenase from Drosophila melanogaster . Biochem Genet 21, 725–733 (1983). https://doi.org/10.1007/BF00498919
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DOI: https://doi.org/10.1007/BF00498919