Skip to main content
NCBI home page
American Journal of Human Genetics logoLink to American Journal of Human Genetics
. 1991 Oct;49(4):878–885.

Homozygosity for a newly identified missense mutation in a patient with very severe combined immunodeficiency due to adenosine deaminase deficiency (ADA-SCID).

R Hirschhorn 1, V Chakravarti 1, J Puck 1, S D Douglas 1
PMCID: PMC1683191  PMID: 1680289

Abstract

We have identified a previously unrecognized missense mutation in a patient with severe combined immunodeficiency due to adenosine deaminase deficiency (ADA-SCID). The mutation is a G646-to-A transition at a CG dinucleotide and predicts a glycine-to-arginine substitution at codon 216. Computer analysis of secondary structure predicts a major alteration with loss of a beta-pleated sheet in a highly conserved region of the protein. The basepair substitution also generates a new site for the restriction enzyme BstXI in exon 7 of the genomic DNA. Digestion of genomic DNA from the patient and from his parents revealed that he was homozygous for the mutation and that his mother and father were carriers. This mutation in homozygous form appears to be associated with very severe disease, since the patient had perinatal onset of clinical manifestations of SCID, the highest concentration of the toxic metabolite deoxyATP in nine patients studied, and a relatively poor immunologic response during the initial 2 years of therapy with polyethylene glycol-adenosine deaminase. Analysis of DNA from 21 additional patients with ADA-SCID and from 19 unrelated normals revealed that, while none of the normal individuals showed the abnormal restriction fragment, two of the 21 patients studied were heterozygous for the G646-to-A mutation.

Full text

PDF
878

Images in this article

883Figure 2
on p.883

Selected References

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

  1. Akeson A. L., Wiginton D. A., Dusing M. R., States J. C., Hutton J. J. Mutant human adenosine deaminase alleles and their expression by transfection into fibroblasts. J Biol Chem. 1988 Nov 5;263(31):16291–16296. [PubMed] [Google Scholar]
  2. Akeson A. L., Wiginton D. A., States J. C., Perme C. M., Dusing M. R., Hutton J. J. Mutations in the human adenosine deaminase gene that affect protein structure and RNA splicing. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5947–5951. doi: 10.1073/pnas.84.16.5947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berkvens T. M., Gerritsen E. J., Oldenburg M., Breukel C., Wijnen J. T., van Ormondt H., Vossen J. M., van der Eb A. J., Meera Khan P. Severe combined immune deficiency due to a homozygous 3.2-kb deletion spanning the promoter and first exon of the adenosine deaminase gene. Nucleic Acids Res. 1987 Nov 25;15(22):9365–9378. doi: 10.1093/nar/15.22.9365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berkvens T. M., van Ormondt H., Gerritsen E. J., Khan P. M., van der Eb A. J. Identical 3250-bp deletion between two AluI repeats in the ADA genes of unrelated ADA-SCID patients. Genomics. 1990 Aug;7(4):486–490. doi: 10.1016/0888-7543(90)90190-6. [DOI] [PubMed] [Google Scholar]
  5. Bonthron D. T., Markham A. F., Ginsburg D., Orkin S. H. Identification of a point mutation in the adenosine deaminase gene responsible for immunodeficiency. J Clin Invest. 1985 Aug;76(2):894–897. doi: 10.1172/JCI112050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cederbaum S. D., Kaitila I., Rimoin D. L., Stiehm E. R. The chondro-osseous dysplasia of adenosine deaminase deficiency with severe combined immunodeficiency. J Pediatr. 1976 Nov;89(5):737–742. doi: 10.1016/s0022-3476(76)80793-7. [DOI] [PubMed] [Google Scholar]
  7. Chang Z. Y., Nygaard P., Chinault A. C., Kellems R. E. Deduced amino acid sequence of Escherichia coli adenosine deaminase reveals evolutionarily conserved amino acid residues: implications for catalytic function. Biochemistry. 1991 Feb 26;30(8):2273–2280. doi: 10.1021/bi00222a033. [DOI] [PubMed] [Google Scholar]
  8. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  9. Cooper D. N., Krawczak M. The mutational spectrum of single base-pair substitutions causing human genetic disease: patterns and predictions. Hum Genet. 1990 Jun;85(1):55–74. doi: 10.1007/BF00276326. [DOI] [PubMed] [Google Scholar]
  10. Daddona P. E., Shewach D. S., Kelley W. N., Argos P., Markham A. F., Orkin S. H. Human adenosine deaminase. cDNA and complete primary amino acid sequence. J Biol Chem. 1984 Oct 10;259(19):12101–12106. [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  12. Garty B. Z., Levinson A. I., Danon Y. L., Wilmott R., Douglas S. D. Lymphocyte subpopulations in children with abnormal lymphatic circulation. J Allergy Clin Immunol. 1989 Oct;84(4 Pt 1):515–520. doi: 10.1016/0091-6749(89)90365-5. [DOI] [PubMed] [Google Scholar]
  13. Giblett E. R., Anderson J. E., Cohen F., Pollara B., Meuwissen H. J. Adenosine-deaminase deficiency in two patients with severely impaired cellular immunity. Lancet. 1972 Nov 18;2(7786):1067–1069. doi: 10.1016/s0140-6736(72)92345-8. [DOI] [PubMed] [Google Scholar]
  14. Hershfield M. S., Buckley R. H., Greenberg M. L., Melton A. L., Schiff R., Hatem C., Kurtzberg J., Markert M. L., Kobayashi R. H., Kobayashi A. L. Treatment of adenosine deaminase deficiency with polyethylene glycol-modified adenosine deaminase. N Engl J Med. 1987 Mar 5;316(10):589–596. doi: 10.1056/NEJM198703053161005. [DOI] [PubMed] [Google Scholar]
  15. Hirschhorn R. Inherited enzyme deficiencies and immunodeficiency: adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) deficiencies. Clin Immunol Immunopathol. 1986 Jul;40(1):157–165. doi: 10.1016/0090-1229(86)90081-4. [DOI] [PubMed] [Google Scholar]
  16. Hirschhorn R., Paageorgiou P. S., Kesarwala H. H., Taft L. T. Amerioration of neurologic abnormalities after "enzyme replacement" in adenosine deaminase deficiency. N Engl J Med. 1980 Aug 14;303(7):377–380. doi: 10.1056/NEJM198008143030706. [DOI] [PubMed] [Google Scholar]
  17. Hirschhorn R., Roegner V., Jenkins T., Seaman C., Piomelli S., Borkowsky W. Erythrocyte adenosine deaminase deficiency without immunodeficiency. Evidence for an unstable mutant enzyme. J Clin Invest. 1979 Oct;64(4):1130–1139. doi: 10.1172/JCI109552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hirschhorn R., Roegner V., Rubinstein A., Papageorgiou P. Plasma deoxyadenosine, adenosine, and erythrocyte deoxyATP are elevated at birth in an adenosine deaminase-deficient child. J Clin Invest. 1980 Mar;65(3):768–771. doi: 10.1172/JCI109725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hirschhorn R., Tzall S., Ellenbogen A. Hot spot mutations in adenosine deaminase deficiency. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6171–6175. doi: 10.1073/pnas.87.16.6171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirschhorn R., Tzall S., Ellenbogen A., Orkin S. H. Identification of a point mutation resulting in a heat-labile adenosine deaminase (ADA) in two unrelated children with partial ADA deficiency. J Clin Invest. 1989 Feb;83(2):497–501. doi: 10.1172/JCI113909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Markert M. L., Hershfield M. S., Wiginton D. A., States J. C., Ward F. E., Bigner S. H., Buckley R. H., Kaufman R. E., Hutton J. J. Identification of a deletion in the adenosine deaminase gene in a child with severe combined immunodeficiency. J Immunol. 1987 May 15;138(10):3203–3206. [PubMed] [Google Scholar]
  22. Markert M. L., Hutton J. J., Wiginton D. A., States J. C., Kaufman R. E. Adenosine deaminase (ADA) deficiency due to deletion of the ADA gene promoter and first exon by homologous recombination between two Alu elements. J Clin Invest. 1988 May;81(5):1323–1327. doi: 10.1172/JCI113458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Markert M. L., Norby-Slycord C., Ward F. E. A high proportion of ADA point mutations associated with a specific alanine-to-valine substitution. Am J Hum Genet. 1989 Sep;45(3):354–361. [PMC free article] [PubMed] [Google Scholar]
  24. Mierendorf R. C., Pfeffer D. Direct sequencing of denatured plasmid DNA. Methods Enzymol. 1987;152:556–562. doi: 10.1016/0076-6879(87)52061-4. [DOI] [PubMed] [Google Scholar]
  25. Orkin S. H., Daddona P. E., Shewach D. S., Markham A. F., Bruns G. A., Goff S. C., Kelley W. N. Molecular cloning of human adenosine deaminase gene sequences. J Biol Chem. 1983 Nov 10;258(21):12753–12756. [PubMed] [Google Scholar]
  26. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schrader W. P., Pollara B., Meuwissen H. J. Characterization of the residual adenosine deaminating activity in the spleen of a patient with combined immunodeficiency disease and adenosine deaminase deficiency. Proc Natl Acad Sci U S A. 1978 Jan;75(1):446–450. doi: 10.1073/pnas.75.1.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  30. Tzall S., Ellenbogen A., Eng F., Hirschhorn R. Identification and characterization of nine RFLPs at the adenosine deaminase (ADA) locus. Am J Hum Genet. 1989 Jun;44(6):864–875. [PMC free article] [PubMed] [Google Scholar]
  31. Valerio D., Dekker B. M., Duyvesteyn M. G., van der Voorn L., Berkvens T. M., van Ormondt H., van der Eb A. J. One adenosine deaminase allele in a patient with severe combined immunodeficiency contains a point mutation abolishing enzyme activity. EMBO J. 1986 Jan;5(1):113–119. doi: 10.1002/j.1460-2075.1986.tb04184.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Valerio D., Duyvesteyn M. G., Dekker B. M., Weeda G., Berkvens T. M., van der Voorn L., van Ormondt H., van der Eb A. J. Adenosine deaminase: characterization and expression of a gene with a remarkable promoter. EMBO J. 1985 Feb;4(2):437–443. doi: 10.1002/j.1460-2075.1985.tb03648.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Valerio D., Duyvesteyn M. G., Meera Khan P., Geurts van Kessel A., de Waard A., van der Eb A. J. Isolation of cDNA clones for human adenosine deaminase. Gene. 1983 Nov;25(2-3):231–240. doi: 10.1016/0378-1119(83)90227-5. [DOI] [PubMed] [Google Scholar]
  34. Valerio D., McIvor R. S., Williams S. R., Duyvesteyn M. G., van Ormondt H., van der Eb A. J., Martin D. W., Jr Cloning of human adenosine deaminase cDNA and expression in mouse cells. Gene. 1984 Nov;31(1-3):147–153. doi: 10.1016/0378-1119(84)90205-1. [DOI] [PubMed] [Google Scholar]
  35. Wiginton D. A., Adrian G. S., Friedman R. L., Suttle D. P., Hutton J. J. Cloning of cDNA sequences of human adenosine deaminase. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7481–7485. doi: 10.1073/pnas.80.24.7481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wiginton D. A., Adrian G. S., Hutton J. J. Sequence of human adenosine deaminase cDNA including the coding region and a small intron. Nucleic Acids Res. 1984 Mar 12;12(5):2439–2446. doi: 10.1093/nar/12.5.2439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wiginton D. A., Kaplan D. J., States J. C., Akeson A. L., Perme C. M., Bilyk I. J., Vaughn A. J., Lattier D. L., Hutton J. J. Complete sequence and structure of the gene for human adenosine deaminase. Biochemistry. 1986 Dec 16;25(25):8234–8244. doi: 10.1021/bi00373a017. [DOI] [PubMed] [Google Scholar]
  38. Wilson D. K., Rudolph F. B., Quiocho F. A. Atomic structure of adenosine deaminase complexed with a transition-state analog: understanding catalysis and immunodeficiency mutations. Science. 1991 May 31;252(5010):1278–1284. doi: 10.1126/science.1925539. [DOI] [PubMed] [Google Scholar]

Articles from American Journal of Human Genetics are provided here courtesy of American Society of Human Genetics

RESOURCES