Sequence of DNA flanking the exons of the HEXA gene, and identification of mutations in Tay-Sachs disease (original) (raw)

. 1991 Nov;49(5):1041–1054.

Abstract

The rapid identification of mutations causing Tay-Sachs disease requires the capacity to readily screen the regions of the HEXA gene most likely to be affected by mutation. We have sequenced the portions of the introns flanking each of the 14 HEXA exons in order to specify oligonucleotide primers for the PCR-dependent amplification of each exon and splice-junction sequence. The amplified products were analyzed, by electrophoresis in nondenaturing polyacrylamide gels, for the presence of either heteroduplexes, derived from the annealing of normal and mutant DNA strands, or single-strand conformational polymorphisms (SSCP), derived from the renaturation of single-stranded DNA. Five novel mutations from Tay-Sachs disease patients were detected: a 5-bp deletion of TCTCC in IVS-9; a 2-bp deletion of TG in exon 5; G78 to A, giving a stop codon in exon 1; G533 to T in exon 5, producing the third amino acid substitution detected at this site; and G to C at position 1 of IVS-2, expected to produce abnormal splicing. In addition, two mutations, (G1496 to A in exon 13 and a 4-bp insertion in exon 11) that have previously been reported were identified.

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Selected References

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  1. Arpaia E., Dumbrille-Ross A., Maler T., Neote K., Tropak M., Troxel C., Stirling J. L., Pitts J. S., Bapat B., Lamhonwah A. M. Identification of an altered splice site in Ashkenazi Tay-Sachs disease. Nature. 1988 May 5;333(6168):85–86. doi: 10.1038/333085a0. [DOI] [PubMed] [Google Scholar]
  2. Brown C. A., Neote K., Leung A., Gravel R. A., Mahuran D. J. Introduction of the alpha subunit mutation associated with the B1 variant of Tay-Sachs disease into the beta subunit produces a beta-hexosaminidase B without catalytic activity. J Biol Chem. 1989 Dec 25;264(36):21705–21710. [PubMed] [Google Scholar]
  3. Conzelmann E., Sandhoff K. Partial enzyme deficiencies: residual activities and the development of neurological disorders. Dev Neurosci. 1983;6(1):58–71. doi: 10.1159/000112332. [DOI] [PubMed] [Google Scholar]
  4. Gelfand M. S. Statistical analysis of mammalian pre-mRNA splicing sites. Nucleic Acids Res. 1989 Aug 11;17(15):6369–6382. doi: 10.1093/nar/17.15.6369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gravel R. A., Triggs-Raine B. L., Mahuran D. J. Biochemistry and genetics of Tay-Sachs disease. Can J Neurol Sci. 1991 Aug;18(3 Suppl):419–423. doi: 10.1017/s0317167100032583. [DOI] [PubMed] [Google Scholar]
  6. Hechtman P., Kaplan F., Bayleran J., Boulay B., Andermann E., de Braekeleer M., Melançon S., Lambert M., Potier M., Gagné R. More than one mutant allele causes infantile Tay-Sachs disease in French-Canadians. Am J Hum Genet. 1990 Nov;47(5):815–822. [PMC free article] [PubMed] [Google Scholar]
  7. Hoar D. I., Haslam D. B., Starozik D. M. Improved direct molecular diagnosis and rapid fetal sexing. Prenat Diagn. 1984 Jul-Aug;4(4):241–247. doi: 10.1002/pd.1970040402. [DOI] [PubMed] [Google Scholar]
  8. Ikehata H., Akagi T., Kimura H., Akasaka S., Kato T. Spectrum of spontaneous mutations in a cDNA of the human hprt gene integrated in chromosomal DNA. Mol Gen Genet. 1989 Nov;219(3):349–358. doi: 10.1007/BF00259606. [DOI] [PubMed] [Google Scholar]
  9. Kazazian H. H., Jr The thalassemia syndromes: molecular basis and prenatal diagnosis in 1990. Semin Hematol. 1990 Jul;27(3):209–228. [PubMed] [Google Scholar]
  10. Korneluk R. G., Mahuran D. J., Neote K., Klavins M. H., O'Dowd B. F., Tropak M., Willard H. F., Anderson M. J., Lowden J. A., Gravel R. A. Isolation of cDNA clones coding for the alpha-subunit of human beta-hexosaminidase. Extensive homology between the alpha- and beta-subunits and studies on Tay-Sachs disease. J Biol Chem. 1986 Jun 25;261(18):8407–8413. [PubMed] [Google Scholar]
  11. Lau M. M., Neufeld E. F. A frameshift mutation in a patient with Tay-Sachs disease causes premature termination and defective intracellular transport of the alpha-subunit of beta-hexosaminidase. J Biol Chem. 1989 Dec 15;264(35):21376–21380. [PubMed] [Google Scholar]
  12. Mahuran D., Novak A., Lowden J. A. The lysosomal hexosaminidase isozymes. Isozymes Curr Top Biol Med Res. 1985;12:229–288. [PubMed] [Google Scholar]
  13. Meuth M. The structure of mutation in mammalian cells. Biochim Biophys Acta. 1990 Jun 1;1032(1):1–17. doi: 10.1016/0304-419x(90)90009-p. [DOI] [PubMed] [Google Scholar]
  14. Myerowitz R., Costigan F. C. The major defect in Ashkenazi Jews with Tay-Sachs disease is an insertion in the gene for the alpha-chain of beta-hexosaminidase. J Biol Chem. 1988 Dec 15;263(35):18587–18589. [PubMed] [Google Scholar]
  15. Myerowitz R., Hogikyan N. D. A deletion involving Alu sequences in the beta-hexosaminidase alpha-chain gene of French Canadians with Tay-Sachs disease. J Biol Chem. 1987 Nov 15;262(32):15396–15399. [PubMed] [Google Scholar]
  16. Myerowitz R., Piekarz R., Neufeld E. F., Shows T. B., Suzuki K. Human beta-hexosaminidase alpha chain: coding sequence and homology with the beta chain. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7830–7834. doi: 10.1073/pnas.82.23.7830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Myerowitz R. Splice junction mutation in some Ashkenazi Jews with Tay-Sachs disease: evidence against a single defect within this ethnic group. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3955–3959. doi: 10.1073/pnas.85.11.3955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Navon R., Kolodny E. H., Mitsumoto H., Thomas G. H., Proia R. L. Ashkenazi-Jewish and non-Jewish adult GM2 gangliosidosis patients share a common genetic defect. Am J Hum Genet. 1990 Apr;46(4):817–821. [PMC free article] [PubMed] [Google Scholar]
  19. Navon R., Proia R. L. The mutations in Ashkenazi Jews with adult GM2 gangliosidosis, the adult form of Tay-Sachs disease. Science. 1989 Mar 17;243(4897):1471–1474. doi: 10.1126/science.2522679. [DOI] [PubMed] [Google Scholar]
  20. Neufeld E. F. Natural history and inherited disorders of a lysosomal enzyme, beta-hexosaminidase. J Biol Chem. 1989 Jul 5;264(19):10927–10930. [PubMed] [Google Scholar]
  21. Orita M., Suzuki Y., Sekiya T., Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics. 1989 Nov;5(4):874–879. doi: 10.1016/0888-7543(89)90129-8. [DOI] [PubMed] [Google Scholar]
  22. Paw B. H., Kaback M. M., Neufeld E. F. Molecular basis of adult-onset and chronic GM2 gangliosidoses in patients of Ashkenazi Jewish origin: substitution of serine for glycine at position 269 of the alpha-subunit of beta-hexosaminidase. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2413–2417. doi: 10.1073/pnas.86.7.2413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paw B. H., Moskowitz S. M., Uhrhammer N., Wright N., Kaback M. M., Neufeld E. F. Juvenile GM2 gangliosidosis caused by substitution of histidine for arginine at position 499 or 504 of the alpha-subunit of beta-hexosaminidase. J Biol Chem. 1990 Jun 5;265(16):9452–9457. [PubMed] [Google Scholar]
  24. Proia R. L., Soravia E. Organization of the gene encoding the human beta-hexosaminidase alpha-chain. J Biol Chem. 1987 Apr 25;262(12):5677–5681. [PubMed] [Google Scholar]
  25. Rozen R., Fox J., Fenton W. A., Horwich A. L., Rosenberg L. E. Gene deletion and restriction fragment length polymorphisms at the human ornithine transcarbamylase locus. 1985 Feb 28-Mar 6Nature. 313(6005):815–817. doi: 10.1038/313815a0. [DOI] [PubMed] [Google Scholar]
  26. Tanaka A., Ohno K., Sandhoff K., Maire I., Kolodny E. H., Brown A., Suzuki K. GM2-gangliosidosis B1 variant: analysis of beta-hexosaminidase alpha gene abnormalities in seven patients. Am J Hum Genet. 1990 Feb;46(2):329–339. [PMC free article] [PubMed] [Google Scholar]
  27. Tindall K. R., Kunkel T. A. Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry. 1988 Aug 9;27(16):6008–6013. doi: 10.1021/bi00416a027. [DOI] [PubMed] [Google Scholar]
  28. Triggs-Raine B. L., Feigenbaum A. S., Natowicz M., Skomorowski M. A., Schuster S. M., Clarke J. T., Mahuran D. J., Kolodny E. H., Gravel R. A. Screening for carriers of Tay-Sachs disease among Ashkenazi Jews. A comparison of DNA-based and enzyme-based tests. N Engl J Med. 1990 Jul 5;323(1):6–12. doi: 10.1056/NEJM199007053230102. [DOI] [PubMed] [Google Scholar]
  29. Triggs-Raine B. L., Gravel R. A. Diagnostic heteroduplexes: simple detection of carriers of a 4-bp insertion mutation in Tay-Sachs disease. Am J Hum Genet. 1990 Jan;46(1):183–184. [PMC free article] [PubMed] [Google Scholar]
  30. White M. B., Amos J., Hsu J. M., Gerrard B., Finn P., Dean M. A frame-shift mutation in the cystic fibrosis gene. Nature. 1990 Apr 12;344(6267):665–667. doi: 10.1038/344665a0. [DOI] [PubMed] [Google Scholar]