Abstract
Complementation of beta hexosaminidase A (hex A) deficiency was obtained by Sendai virus-mediated somatic cell hybridization of cultured skin fibroblasts from two unrelated patients with Tay-Sachs disease (TSD) and one patient with Sandhoff-Jatzkewitz disease (SJD). The newly formed hex A was identified by its electrophoretic mobility in three different systems, heat lability, and reactivity with an antiserum against the unique antigenic determinant, alpha of hex A. The percentage of heterokaryons obtained by virus treatment of TSD and SJD fibroblast mixtures showed good correlation with the observed percentage of hex A activity. It is concluded that, in these two forms of GM2 gangliosidosis, beta hexosaminidase deficiency results from two different mutations. All of the current models of beta hexosaminidase structure are compatible with the observed complementation. No complementation was detected in 13 Sendai virus-induced fusions of cultured skin fibroblasts from seven unrelated patients with SJD. The enzyme deficiency in these patients may be due to very similar allelic mutations, not capable of undergoing complementation; or to different structural mutations, all coding for unstable beta hexosaminidase molecules.
MeSH Terms
Acetylglucosaminidase/deficiency
Cell Fusion
Cells, Cultured
Female
Fibroblasts/enzymology
Gangliosidoses/enzymology,genetics
Genetic Complementation Test
Hexosaminidases/deficiency
Humans
Lipidoses/genetics
Male
Nucleic Acid Hybridization
Skin/enzymology
Chemicals
Hexosaminidases
Acetylglucosaminidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rattazzi M C
Brown J A
Davidson R G
Shows T B
References (29)
29 references, click to expand
-
Activation of production of infectious tumor virus SV40 in heterokaryon cultures.
Proc Natl Acad Sci U S A. 1967 Jul;58(1):127-33
PMID: 4292099
-
N-Acetyl-beta-glucosaminidases in human spleen.
Biochem J. 1968 Apr;107(3):321-7
PMID: 5650361
-
Lysosomal hydrolases: Conversion of acidic to basic forms by neuraminidase.
FEBS Lett. 1971 Feb 12;13(1):68-72
PMID: 11945635
-
A simple ultraviolet spectrophotometric method for the determination of protein.
J Lab Clin Med. 1956 Aug;48(2):311-4
PMID: 13346201
-
Human beta-D-N-acetylhexosaminidases A and B: expression and linkage relationships in somatic cell hybrids.
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1569-73
PMID: 4524661
-
Hydrolysis of Tay-Sachs ganglioside by beta-hexosaminidase A of human liver and urine.
J Biol Chem. 1973 Nov 10;248(21):7512-5
PMID: 4745777
-
Gm2-gangliosidosis with total hexosaminidase deficiency.
Neurology. 1971 Apr;21(4):313-28
PMID: 4251893
-
Genetic complementation after fusion of Tay-Sachs and Sandhoff cells.
Nature. 1974 Aug 16;250(467):580-2
PMID: 4367631
-
Isolation and relationship of human hexosaminidases.
J Biol Chem. 1974 Jun 10;249(11):3489-99
PMID: 4364658
-
Complementation between alleles in heterocaryons.
Cold Spring Harb Symp Quant Biol. 1958;23:137-40
PMID: 13635550
-
Immunological properties of N-acetyl-beta-D-glucosaminidase of normal human liver and of GM2-gangliosidosis liver.
Biochem J. 1973 Jan;131(1):91-6
PMID: 4198585
-
Studies on human beta-D-N-acetylhexosaminidases. 3. Biochemical genetics of Tay-Sachs and Sandhoff's diseases.
J Biol Chem. 1974 Apr 10;249(7):2054-7
PMID: 4206549
-
Hybrids between human leukocytes and a mouse cell line: production and characterization.
Wistar Inst Symp Monogr. 1969;9:61-76
PMID: 5408740
-
Studies on human beta-D-N-acetylhexosaminidases. I. Purification and properties.
J Biol Chem. 1974 Apr 10;249(7):2043-8
PMID: 4362060
-
Deficient hexosaminidase activity in an exceptional case of Tay-Sachs disease with additional storage of kidney globoside in visceral organs.
Pathol Eur. 1968;3(2):278-85
PMID: 5688464
-
[Enzymatic studies on the blood of carriers of a Tay-Sachs disease variant (variant O)].
Klin Wochenschr. 1971 Nov 1;49(21):1189-91
PMID: 5124584
-
Tay-Sachs and Sandhoff's disease: intergenic complementation after somatic cell hybridization.
Exp Cell Res. 1974 Aug;87(2):444-8
PMID: 4416048
-
Immunochemical characterization of human beta-D-N-acetyl hexosaminidase from normal individuals and patients with Tay-Sachs disease. I. Antigenic differences between hexosaminidase A and hexosaminidase B.
Int Arch Allergy Appl Immunol. 1974;46(4):512-24
PMID: 4132060
-
Sandhoff's disease (GM 2 gangliosidosis type 2): clinical, chemical, and enzyme studies in five patients.
Pediatr Res. 1972 Jul;6(7):606-15
PMID: 5057290
-
Studies on the substrate specificity of hexosaminidase A and B from liver.
Arch Biochem Biophys. 1972 Nov;153(1):116-29
PMID: 4346630
-
Complementation after fusion of Sandhoff- and Tay-Sachs fibroblasts.
Humangenetik. 1975;26(2):117-21
PMID: 803466
-
Enzyme alterations and lipid storage in three variants of Tay-Sachs disease.
J Neurochem. 1971 Dec;18(12):2469-89
PMID: 5135907
-
Ganglioside GM2 storage diseases: hexosaminidase deficiencies in cultured fibroblasts.
Am J Hum Genet. 1971 Jan;23(1):55-61
PMID: 5581981
-
Tay-Sachs disease: generalized absence of a beta-D-N-acetylhexosaminidase component.
Science. 1969 Aug 15;165(3894):698-700
PMID: 5793973
-
[Enrichment and characterization of 2 forms of human N-acetyl-beta-D-hexosaminidase].
Hoppe Seylers Z Physiol Chem. 1971 Aug;352(8):1119-33
PMID: 5098352
-
Studies on human beta-D-N-acetylhexosaminidases. II. Kinetic and structural properties.
J Biol Chem. 1974 Apr 10;249(7):2049-53
PMID: 4818822
-
THE THEORY OF INTER-ALLELIC COMPLEMENTATION.
J Mol Biol. 1964 Jan;8:161-5
PMID: 14149958
-
Characterization of Hex S, the major residual beta hexosaminidase activity in type O Gm2 gangliosidosis (Sandhoff-Jatzkewitz disease).
Am J Hum Genet. 1975 Sep;27(5):639-50
PMID: 240271
-
Studies on glucosaminidase. 4. The fluorimetric assay of N-acetyl-beta-glucosaminidase.
Biochem J. 1961 Jan;78:151-6
PMID: 13759894