Abstract
We have examined genetic complementation in pyruvate carboxylase deficiency by comparing the enzyme activity in polyethylene glycol-induced heterokaryons with that in unfused mixtures of fibroblasts from three affected children. Complementation, manifested as a three- to sevenfold increase in pyruvate carboxylase activity, was observed in fusions between a biotin-responsive multiple carboxylase (pyruvate carboxylase, propionyl CoA carboxylase, and β-methylcrotonyl CoA carboxylase) deficient fibroblast line and two other lines deficient only in pyruvate carboxylase activity. Kinetic analysis of complementing pyruvate carboxylase deficient lines, measured by the rate of restoration of enzyme activity as a function of time, revealed that maximum restoration was achieved within 10–24 hr after fusion. This profile is similar to those observed for fusions between the multiple carboxylase deficient line and two lines deficient in propionyl CoA carboxylase activity that are known to represent different gene mutations. Although the patients with pyruvate carboxylase deficiency had similar clinical findings, our studies indicate that pyruvate carboxylase deficiency is genetically heterogeneous, with at least two distinct, probably intergenic, complementation groups.
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Atkin, B. M., Utter, M. F., and Weinberg, M. B. (1979) Pyruvate carboxylase and phosphoenolpyruvate carboxykinase in leukocytes and fibroblasts from a patient with pyruvate carboxylase deficiency. Pediatr. Res. 1338.
Barden, R. E., Taylor, B. L., Isohashi, F., Frey, W. H., II, Zander, G., Lee, J. C., and Utter, M. F. (1975). Structural properties of pyruvate carboxylase from chicken liver and other sources. Proc. Natl. Acad. Sci. 724308.
Brunette, M. B., Delvin, E., Hazel, B., and Scriver, C. R. (1972). Thiamine-responsive lactic acidosis in a patient with deficient low K m pyruvate carboxylase activity in liver. Pediatrics 50702.
DeVivo, D. C., Haymond, M. W., Leckie, M. P., Bussmann, Y. L., McDougal, Jr., D. B., and Pagliara, A. S. (1977). The clinical and biochemical implications of pyruvate carboxylase deficiency. J. Clin. Endocrinol. Metab. 451281.
Gravel, R. A., Lam, F.-K., Scully, K. J., and Hsia, Y. E. (1977). Genetic complementation of propionyl CoA carboxylase deficiency in cultured fibroblasts. Am. J. Hum. Genet. 29378.
Grobe, H., v. Bassiwitz, D. B., Dominick, H.-Chr., and Pfeiffer, R. A. (1975). Subacute necrotizing encephalomyelopathy: Clinical, ultrastructural, biochemical and therapeutic studies in an infant. Acta Paediatr. Scand. 64755.
Grover, W. D., Auerbach, V. H., and Patel, M. S. (1972). Biochemical studies and therapy in subacute necrotizing encephalomyelopathy (Leigh's syndrome). J. Pediatr. 8139.
Gruskin, A. B., Patel, M. S., Linshaw, M., Ettenger, R., Huff, D., and Grover, W. (1973). Renal function studies and kidney pyruvate carboxylase in subacute necrotizing encephalomyelopathy (Leigh's syndrome). Pediatr. Res. 7832.
Hommes, F. A., Polman, H. A., and Reerink, J. D. (1968). Leigh's encephalomyelopathy: An inborn error of gluconeogenesis. Arch. Dis. Childh. 43423.
Kalousek, F., Darigo, M. D., and Rosenberg, L. E. (1979). Structural and functional features of homogeneous propionyl CoA carboxylase (PCC) from human liver. XIth Int. Congr. Biochem., p. 276.
Kornberg, H. L. (1966). Campbell, P. N., and Greville, G. D. (ed.), Essays in Biochemistry, Vol. 2, Academic Press, New York, p. 131.
Lowry, D. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the folin phenol reagent. J. Biol. Chem. 193265.
Maesaka, H., Komiya, K., Misugi, K., and Tada, K. (1976). Hyperalaninemia, hyperpyruvicemia and lactic acidosis due to pyruvate carboxylase deficiency in the liver; treatment with thiamine and lipoic acid. Eur. J. Pediatr. 122159.
Roth, K., Cohn, R., Yandrasitz, J., Preti, G., Dodd, P., and Segal, S. (1976). Beta-methylcrotonic aciduria associated with lactic acidosis. J. Pediatr. 88229.
Saudubray, J. M., Marsac, C., Charpentier, C., Cathelineau, L., Leaud, M. B., and Leroux, J. P. (1976). Neonatal congenital lactic acidosis with pyruvate carboxylase deficiency in two sublings. Acta Paediatr. Scand. 65717.
Scrutton, M. C., and White, M. D. (1974). Purification and properties of human liver pyruvate carboxylase. Biochem. Med. 9271.
Sutcliffe, R. G., Carritt, B., and Wilson, R. H. (1978). Genes, proteins, and the control of gene expression. In (eds.), Brock D. J. H., and Mayo, O., The Biochemical Genetics of Man, Academic Press, New York, pp. 42–45.
Sweetman, L., Packman, S., Yoshino, M., Cowan, M., Wara, D., Ammann, A., and Nyhan, W. (1979). Biotin responsive multiple carboxylase deficiency. Pediatr. Res. 13602.
Tada, K., Sugita, K., Fujitani, K., Uesakai, T., Takada, G., and Omura, K. (1973). Hyperalaninemia with pyruvicemia in a patient suggestive of Leigh's encephalomyelopathy. Tohoku J. Exp. Med. 10913.
Tang, T. T., Good, T. A., Dyken, P. R., Johnsen, S. D., McCreadie, S. R., Sy, S. T., Lardy, H. A., and Rudolph, F. B. (1972). Pathogenesis of Leigh's encephalomyelopathy. J. Pediatr. 81189.
Utter, M. F., and Keech, D. B. (1963). Pyruvate carboxylase I. Nature of the reaction. J. Biol. Chem. 2382603.
Weyler, W., Sweetman, L., Maggio, D. C., and Nyhan, W. L. (1977). Deficiency of propionyl CoA carboxylase and β-methylcrotonyl CoA carboxylase in a patient with methylcrotonylglycinuria. Clin. Chim. Acta 76321.
Willard, H. F., Mellman, I. S., and Rosenberg, L. E. (1978). Genetic complementation among inherited deficiencies of methylmalonyl-CoA mutase activity: Evidence for a new class of human cobalamin mutant. Am. J. Hum. Genet. 301.
Wolf, B., and Rosenberg, L. E. (1978). Heterozygote expression in propionyl coenzyme A carboxylase deficiency. J. Clin. Invest. 62931.
Wolf, B., Hsia, Y. E., and Rosenberg, L. E. (1978). Biochemical differences between mutant propionyl CoA carboxylases from two complementation groups. Am. J. Hum. Genet. 30455.
Wolf, B., Willard, H. F., and Rosenberg, L. E. (1980). Kinetic analysis of genetic complementation in heterokaryons of propionyl CoA carboxylase deficient human fibroblasts. Am. J. Hum. Genet. 3216.
Yoshida, T., Tada, K., Konno, T., and Arakawa, T. (1969). Hyperalaninemia with pyruvicemia due to pyruvate carboxylase deficiency of the liver. Tohoku J. Exp. Med. 99121.
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This work was supported by an NIH research grant (AM 25675) and an A. D. Williams research grant (6-48360). B. Wolf is the recipient of an NIH Research Career Development Award (AM 00677) and is aided by a Basil O'Connor Starter Research Grant from The National Foundation-March of Dimes (5-263). G. Feldman is the recipient of an NIH predoctoral training grant (GM 07492). This article is No. 100 from the Department of Human Genetics at the Medical College of Virginia.
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Feldman, G.L., Wolf, B. Evidence for two genetic complementation groups in pyruvate carboxylase-deficient human fibroblast cell lines. Biochem Genet 18, 617–624 (1980). https://doi.org/10.1007/BF00484405
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DOI: https://doi.org/10.1007/BF00484405