Summary
Cell hybridizations between fibroblasts of four variants (B, O, AB, and B1) of infantile GM2 gangliosidosis were performed. Cocultivated as well as hybrid cells were analyzed for their capability to degrade exogenously added [3H]-GM2. Hybridization of variant AB fibroblasts with fibroblasts of variant O, variant B, or variant B1 resulted in an enhanced rate of GM2 hydrolysis, showing intergenic complementation. Similar restoration of GM2 catabolism was observed after hybridization of variant B1 cells with variant O, but not with variant B cells. These results indicate that B1 cells carry a mutation in the gene locus for the α-subunit of β-hexosaminidase. Studies of the processing of immature enzyme in variant B1 cells showed the presence of α-precursors and mature α-chains, but at a lower level as compared to normal cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bonner WM, Laskey RA (1974) A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46:83–88
Burg J (1984) Immunochemische Charakterisierung des Gangliosid GM2-Aktivatorproteins: Biosynthese und Verteilung in normalen und pathologischen Geweben. Dissertation, Universität Bonn
Conzelmann E, Sandhoff K (1978) AB variant of infantile GM2 gangliosidosis: deficiency of a factor necessary for stimulation of hexosaminidase A-catalyzed degradation of ganglioside GM2 and glycolipid GA2. Proc Natl Acad Sci USA 75:3979–3983
Galjaard H, Hoogeveen A, de Wit-Verbeek HA, Reuser AJJ, Keijzer W, Westerveld A, Bottsma D (1974) Tay-Sachs and Sandhoff disease: intergenic complementation after somatic cell hybridization. Exp Cell Res 87:444–448
Goldman JE, Yamanaka T, Rapin I, Adachi M, Suzuki K, Suzuki K (1980) The AB-variant of GM2-gangliosidosis. Clinical, biochemical and pathological studies of two patients. Acta Neuropathol (Berl) 52:189–202
Hasilik A, Neufeld EF (1980) Biosynthesis of lysosomal enzymes in fibroblasts: synthesis as precursors of higher molecular weight. J Biol Chem 255:4937–4945
Hirabayashi Y, Li YT, Li SC (1983) The protein activator specific for the enzymic hydrolysis of GM2 ganglioside in normal human brain and brains of three types of GM2 gangliosidosis. J Neurochem 40:168–175
Hoogeveen AT, Verheijen FW, D'Azzo A, Galjaard H (1980) Genetic heterogeneity in human neuraminidase deficiency. Nature 285:500–502
Kytzia HJ, Hinrichs U, Maire I, Suzuki K, Sandhoff K (1983) Variant of GM2-gangliosidosis with hexosaminidase A having a severely changed substrate specificity. EMBO J 2:1201–1205
Kytzia HJ, Hinrichs U, Sandhoff K (1984) Diagnosis of infantile and juvenile forms of GM2 gangliosidosis variant O. Residual activities toward natural and different synthetic substrates. Hum Genet 67:414–418
Kytzia HJ, Sandhoff K (1985) Evidence for two different active sites on human β-hexosaminidase A. Interaction of GM2 activator protein with β-hexosaminidase A. J Biol Chem 260:7568–7572
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature 227:680–685
Laskey RA, Mills AD (1975) Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem 56:335–341
Li SC, Hirabayashi Y, Li YT (1981) A new variant of type AB GM2-gangliosidosis. Biochem Biophys Res Commun 101:479–485
Myerowitz R, Proia RL (1984) CDNA clone for the α chain of human β-hexosaminidase: deficiency of α-chain mRNA in Ashkenazi Tay-Sachs fibroblasts. Proc Natl Acad Sci USA 81:5394–5398
Neufeld EF (1981) Recognition and processing of lysosomal enzymes in cultured fibroblasts. In: Callahan JW, Lowden JA (eds) Lysosomes and lysosomal storage diseases. Raven Press, New York, pp 115–130
O'Brien JS (1983) The gangliosidosis. In: Stanbury J, Wyngaarden J, Fredrickson D, Goldstein J, Brown M (eds) The metabolic basis of inherited disease. McGraw-Hill, New York, pp 945–969
Okada S, O'Brien JS (1969) Tay-Sachs disease: generalized absence of a beta-D-N-acetylhexosaminidase component. Science 165: 698–700
Proia RL, D'Azzo A, Neufeld EF (1984) Association of α- and β-subunits during the biosynthesis of β-hexosaminidase in cultured human fibroblasts. J Biol Chem 259:3350–3354
Rattazzi MC, Brown JA, Davidson RG, Shows TB (1975) Tay-Sachs and Sandhoff-Jatzkewitz diseases: complementation of hexosaminidase A deficiency by somatic cell hybridization. Cytogenet Cell Genet 14:232–235
Ropers HH, Grezschik KH, Bühler E (1975) Complementation after fusion of Sandhoff- and Tay-Sachs-fibroblasts. Humangenetik 26:116–121
Sandhoff K (1969) Variation of β-N-acetylhexosaminidase-pattern in Tay-Sachs disease. FEBS Lett 4:351–354
Sandhoff K (1984) Function and relevance of activator proteins for glycolipid degradation. In: Brady RO, Barranger JA (eds) Molecular basis of lysosomal storage disorders. Academic Press, New York, pp 19–49
Sandhoff K, Christomanou H (1979) Biochemistry and genetics of gangliosidoses. Hum Genet 50:107–143
Schwarzmann G (1978) A simple and novel method for tritium labeling of gangliosides and other sphingolipids. Biochim Biophys Acta 529:106–114
Sonderfeld S, Conzelmann E, Schwarzmann G, Burg J, Hinrichs U, Sandhoff K (1985) Incorporation and metabolism of ganglioside GM2 in normal and GM2 gangliosidosis skin fibroblasts. Eur J Biochem 149:247–255
Thomas GH, Taylor HA, Miller CS, Axelman J, Migeon BR (1974) Genetic complementation after fusion of Tay-Sachs and Sandhoff cells. Nature 250:580–582
Weitz G, Lindl T, Hinrichs U, Sandhoff K (1983) Release of sphingomyelin phosphodiesterase (acid sphingomyelinase) by ammonium chloride from Cl 1 D mouse L-cells and human fibroblasts. Hoppe-Seyler's Z Physiol Chem 364:863–871
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sonderfeld, S., Brendler, S., Sandhoff, K. et al. Genetic complementation in somatic cell hybrids of four variants of infantile GM2 gangliosidosis. Hum Genet 71, 196–200 (1985). https://doi.org/10.1007/BF00284572
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00284572