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. 1974 Apr;71(4):1569–1573. doi: 10.1073/pnas.71.4.1569

Human β-D-N-Acetylhexosaminidases A and B: Expression and Linkage Relationships in Somatic Cell Hybrids

P A Lalley *, M C Rattazzi †, T B Shows *
PMCID: PMC388272  PMID: 4524661

Abstract

Knowledge of the genetic relationships between β-D-N-acetylhexosaminidases A and B (EC 3.2.1.30) may help in understanding the hexosaminidase deficiency associated with GM2 gangliosidosis, a fatal lipid storage disease in man. Through the use of man-mouse somatic cell hybrids we have found that a gene involved in hexosaminidase A expression was linked to the genes coding for mannosephosphate isomerase and pyruvate kinase-3. The gene coding for hexosaminidase B was not linked to any of the genes coding for 25 enzyme markers tested. A combination of immunological and electrophoretic techniques was employed to identify human hexosaminidases A and B with certainty in cell hybrids. Discordant segregation of hexosaminidase A and hexosaminidase B in 60 clones indicated that the genes coding for their expression were not linked. However, hexosaminidase A was never expressed in cell hybrids in the absence of hexosaminidase B. This suggests that the gene responsible for the hexosaminidase A phenotype, linked to mannosephosphate isomerase and pyruvate kinase-3, requires the presence of the gene coding for hexosaminidase B for the expression of hexosaminidase A. These observations offer a genetic explanation for the biochemical and immunological relationships between hexosaminidases A and B and provide the framework for identifying the basic genetic defects responsible for GM2 gangliosidosis.

Keywords: GM2 gangliosidosis, lysosomal enzymes, anti-hexosaminidase sera, electrophoresis

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bartholomew W. R., Rattazzi M. C. 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):512–524. doi: 10.1159/000231154. [DOI] [PubMed] [Google Scholar]
  2. Carroll M., Robinson D. Immunological properties of N-acetyl-beta-D-glucosaminidase of normal human liver and of GM2-gangliosidosis liver. Biochem J. 1973 Jan;131(1):91–96. doi: 10.1042/bj1310091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen S. H., Malcolm L. A., Yoshida A., Giblett E. R. Phosphoglycerate kinase: an X-linked polymorphism in man. Am J Hum Genet. 1971 Jan;23(1):87–91. [PMC free article] [PubMed] [Google Scholar]
  4. Cohen M. M., Lin C. C., Sybert V., Orecchio E. J. Two human X-autosome translocations identified by autoradiography and fluorescence. Am J Hum Genet. 1972 Sep;24(5):583–597. [PMC free article] [PubMed] [Google Scholar]
  5. Edwards Y. H., Hopkinson D. A., Harris H. Inherited variants of human nucleoside phosphorylase. Ann Hum Genet. 1971 May;34(4):395–408. doi: 10.1111/j.1469-1809.1971.tb00252.x. [DOI] [PubMed] [Google Scholar]
  6. Egli F., Stalder G. Malformations of kidney and urinary tract in common chromosomal aberrations. I. Clinical studies. Humangenetik. 1973 Mar 23;18(1):1–15. doi: 10.1007/BF00279026. [DOI] [PubMed] [Google Scholar]
  7. Goldstone A., Konecny P., Koenig H. Lysosomal hydrolases: Conversion of acidic to basic forms by neuraminidase. FEBS Lett. 1971 Feb 12;13(1):68–72. doi: 10.1016/0014-5793(71)80667-1. [DOI] [PubMed] [Google Scholar]
  8. HAYFLICK L., MOORHEAD P. S. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961 Dec;25:585–621. doi: 10.1016/0014-4827(61)90192-6. [DOI] [PubMed] [Google Scholar]
  9. Klebe R. J., Chen T., Ruddle F. H. Controlled production of proliferating somatic cell hybrids. J Cell Biol. 1970 Apr;45(1):74–82. doi: 10.1083/jcb.45.1.74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Klebe R. J., Chen T., Ruddle R. H. Mapping of a human genetic regulator element by somatic cell genetic analysis. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1220–1227. doi: 10.1073/pnas.66.4.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LEABACK D. H., WALKER P. G. Studies on glucosaminidase. 4. The fluorimetric assay of N-acetyl-beta-glucosaminidase. Biochem J. 1961 Jan;78:151–156. doi: 10.1042/bj0780151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  13. McMorris F. A., Chen T. R., Ricciuti F., Tischfield J., Creagan R., Ruddle F. Chromosome assignments in man of the genes for two hexosephosphate isomerases. Science. 1973 Mar 16;179(4078):1129–1131. doi: 10.1126/science.179.4078.1129. [DOI] [PubMed] [Google Scholar]
  14. Mowbray S., Watson B., Harris H. A search for electrophoretic variants of human adenine phosphoribosyl transferase. Ann Hum Genet. 1972 Nov;36(2):153–162. doi: 10.1111/j.1469-1809.1972.tb00766.x. [DOI] [PubMed] [Google Scholar]
  15. Nichols E. A., Chapman V. M., Ruddle F. H. Polymorphism and linkage for mannosephosphate isomerase in Mus musculus. Biochem Genet. 1973 Jan;8(1):47–53. doi: 10.1007/BF00485556. [DOI] [PubMed] [Google Scholar]
  16. Okada S., O'Brien J. S. Tay-Sachs disease: generalized absence of a beta-D-N-acetylhexosaminidase component. Science. 1969 Aug 15;165(3894):698–700. doi: 10.1126/science.165.3894.698. [DOI] [PubMed] [Google Scholar]
  17. PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ritter H., Schmitt J. Genetic polymorphism of mannosephosphate isomerase in Primates. Humangenetik. 1973 Sep 20;19(3):325–326. doi: 10.1007/BF00278411. [DOI] [PubMed] [Google Scholar]
  19. Robinson D., Stirling J. L. N-Acetyl-beta-glucosaminidases in human spleen. Biochem J. 1968 Apr;107(3):321–327. doi: 10.1042/bj1070321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ruddle F. H. Linkage analysis in man by somatic cell genetics. Nature. 1973 Mar 16;242(5394):165–169. doi: 10.1038/242165a0. [DOI] [PubMed] [Google Scholar]
  21. Sandhoff K., Harzer K., Wässle W., Jatzkewitz H. Enzyme alterations and lipid storage in three variants of Tay-Sachs disease. J Neurochem. 1971 Dec;18(12):2469–2489. doi: 10.1111/j.1471-4159.1971.tb00204.x. [DOI] [PubMed] [Google Scholar]
  22. Shin S., Meera Khan P., Cook P. R. Characterization of hypoxanthine-guanine phosphoribosyl transferase in man--mouse somatic cell hybrids by an improved electrophoretic method. Biochem Genet. 1971 Feb;5(1):91–99. doi: 10.1007/BF00485734. [DOI] [PubMed] [Google Scholar]
  23. Shows T. B. Genetics of human-mouse somatic cell hybrids: linkage of human genes for isocitrate dehydrogenase and malate dehydrogenase. Biochem Genet. 1972 Dec;7(3):193–204. doi: 10.1007/BF00484817. [DOI] [PubMed] [Google Scholar]
  24. Shows T. B. Genetics of human-mouse somatic cell hybrids: linkage of human genes for lactate dehydrogenase-A and esterase-A 4 . Proc Natl Acad Sci U S A. 1972 Feb;69(2):348–352. doi: 10.1073/pnas.69.2.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shows T. B., Ruddle F. H. Function of the lactate dehydrogenase B gene in mouse erythrocytes: evidence for control by a regulatory gene. Proc Natl Acad Sci U S A. 1968 Oct;61(2):574–581. doi: 10.1073/pnas.61.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Srivastava S. K., Beutler E. Hexosaminidase-A and hexosaminidase-B: studies in Tay-Sachs' and Sandhoff's disease. Nature. 1973 Feb 16;241(5390):463–463. doi: 10.1038/241463a0. [DOI] [PubMed] [Google Scholar]
  27. Suzuki Y., Suzuki K. Partial deficiency of hexosaminidase component a in juvenile gm2-gangliosidosis. Neurology. 1970 Sep;20(9):848–851. doi: 10.1212/wnl.20.9.848. [DOI] [PubMed] [Google Scholar]
  28. Swallow D. M., Stokes D. C., Corney G., Harris H. Differences between the N-acetyl hexosaminidase isozymes in serum and tissues. Ann Hum Genet. 1974 Jan;37(3):287–302. doi: 10.1111/j.1469-1809.1974.tb01836.x. [DOI] [PubMed] [Google Scholar]

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