A single-base-pair deletion in the beta-glucuronidase gene accounts for the phenotype of murine mucopolysaccharidosis type VII (original) (raw)

Abstract

Murine mucopolysaccharidosis type VII is a heritable disease caused by a spontaneous mutation, gus(mps), closely linked to the beta-glucuronidase structural gene on chromosome 5. Mice homozygous for the mutation have a > 200-fold decrease in beta-glucuronidase mRNA levels and virtually no enzyme activity detectable by a sensitive fluorometric assay. Approximately 20 kb of genomic DNA containing the beta-glucuronidase gene Gus and > 2 kb of 5' and 3' flanking sequences were cloned from both a gus(mps)/gus(mps) mouse and a +/+ mouse of the progenitor strain. Restriction enzyme digests containing DNA fragments 20-400 bp in length were generated from each of the two Gus alleles and then compared by using nondenaturing polyacrylamide DNA-sequencing gels. This method rapidly identified a large number of restriction sites and was sensitive enough to detect a restriction fragment length variation resulting from a 1-bp deletion in the gus(mps) allele. DNA-sequence analysis of the mutant genomic fragment showed that the 1-bp deletion created a frameshift mutation within exon 10. Insertion of the deleted nucleotide by oligonucleotide site-directed mutagenesis restored function to the corrected mutant gene when transfected into gus(mps)/gus(mps) fibroblasts. We concluded that the frameshift mutation, which introduces a premature stop codon at codon 497 in exon 10, accounts for the molecular, biochemical, and pathological abnormalities associated with the gus(mps) phenotype.

6567

Images in this article

Selected References

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

  1. Baserga S. J., Benz E. J., Jr Beta-globin nonsense mutation: deficient accumulation of mRNA occurs despite normal cytoplasmic stability. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2935–2939. doi: 10.1073/pnas.89.7.2935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birkenmeier E. H., Davisson M. T., Beamer W. G., Ganschow R. E., Vogler C. A., Gwynn B., Lyford K. A., Maltais L. M., Wawrzyniak C. J. Murine mucopolysaccharidosis type VII. Characterization of a mouse with beta-glucuronidase deficiency. J Clin Invest. 1989 Apr;83(4):1258–1266. doi: 10.1172/JCI114010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boguski M. S., Birkenmeier E. H., Elshourbagy N. A., Taylor J. M., Gordon J. I. Evolution of the apolipoproteins. Structure of the rat apo-A-IV gene and its relationship to the human genes for apo-A-I, C-III, and E. J Biol Chem. 1986 May 15;261(14):6398–6407. [PubMed] [Google Scholar]
  4. Cheng J., Fogel-Petrovic M., Maquat L. E. Translation to near the distal end of the penultimate exon is required for normal levels of spliced triosephosphate isomerase mRNA. Mol Cell Biol. 1990 Oct;10(10):5215–5225. doi: 10.1128/mcb.10.10.5215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. D'Amore M. A., Gallagher P. M., Korfhagen T. R., Ganschow R. E. Complete sequence and organization of the murine beta-glucuronidase gene. Biochemistry. 1988 Sep 6;27(18):7131–7140. doi: 10.1021/bi00418a070. [DOI] [PubMed] [Google Scholar]
  6. Dietz H. C., Valle D., Francomano C. A., Kendzior R. J., Jr, Pyeritz R. E., Cutting G. R. The skipping of constitutive exons in vivo induced by nonsense mutations. Science. 1993 Jan 29;259(5095):680–683. doi: 10.1126/science.8430317. [DOI] [PubMed] [Google Scholar]
  7. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kyle J. W., Birkenmeier E. H., Gwynn B., Vogler C., Hoppe P. C., Hoffmann J. W., Sly W. S. Correction of murine mucopolysaccharidosis VII by a human beta-glucuronidase transgene. Proc Natl Acad Sci U S A. 1990 May;87(10):3914–3918. doi: 10.1073/pnas.87.10.3914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mashima Y., Murakami A., Weleber R. G., Kennaway N. G., Clarke L., Shiono T., Inana G. Nonsense-codon mutations of the ornithine aminotransferase gene with decreased levels of mutant mRNA in gyrate atrophy. Am J Hum Genet. 1992 Jul;51(1):81–91. [PMC free article] [PubMed] [Google Scholar]
  10. Naeger L. K., Schoborg R. V., Zhao Q., Tullis G. E., Pintel D. J. Nonsense mutations inhibit splicing of MVM RNA in cis when they interrupt the reading frame of either exon of the final spliced product. Genes Dev. 1992 Jun;6(6):1107–1119. doi: 10.1101/gad.6.6.1107. [DOI] [PubMed] [Google Scholar]
  11. Paigen K. Mammalian beta-glucuronidase: genetics, molecular biology, and cell biology. Prog Nucleic Acid Res Mol Biol. 1989;37:155–205. doi: 10.1016/s0079-6603(08)60698-4. [DOI] [PubMed] [Google Scholar]
  12. Powell P. P., Kyle J. W., Miller R. D., Pantano J., Grubb J. H., Sly W. S. Rat liver beta-glucuronidase. cDNA cloning, sequence comparisons and expression of a chimeric protein in COS cells. Biochem J. 1988 Mar 1;250(2):547–555. doi: 10.1042/bj2500547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Shapiro M. B., Senapathy P. RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res. 1987 Sep 11;15(17):7155–7174. doi: 10.1093/nar/15.17.7155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sly W. S., Quinton B. A., McAlister W. H., Rimoin D. L. Beta glucuronidase deficiency: report of clinical, radiologic, and biochemical features of a new mucopolysaccharidosis. J Pediatr. 1973 Feb;82(2):249–257. doi: 10.1016/s0022-3476(73)80162-3. [DOI] [PubMed] [Google Scholar]
  17. Urlaub G., Mitchell P. J., Ciudad C. J., Chasin L. A. Nonsense mutations in the dihydrofolate reductase gene affect RNA processing. Mol Cell Biol. 1989 Jul;9(7):2868–2880. doi: 10.1128/mcb.9.7.2868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Vogler C., Birkenmeier E. H., Sly W. S., Levy B., Pegors C., Kyle J. W., Beamer W. G. A murine model of mucopolysaccharidosis VII. Gross and microscopic findings in beta-glucuronidase-deficient mice. Am J Pathol. 1990 Jan;136(1):207–217. [PMC free article] [PubMed] [Google Scholar]
  19. Watakabe A., Tanaka K., Shimura Y. The role of exon sequences in splice site selection. Genes Dev. 1993 Mar;7(3):407–418. doi: 10.1101/gad.7.3.407. [DOI] [PubMed] [Google Scholar]
  20. Wolfe J. H., Sands M. S., Barker J. E., Gwynn B., Rowe L. B., Vogler C. A., Birkenmeier E. H. Reversal of pathology in murine mucopolysaccharidosis type VII by somatic cell gene transfer. Nature. 1992 Dec 24;360(6406):749–753. doi: 10.1038/360749a0. [DOI] [PubMed] [Google Scholar]
  21. Wolfe J. H., Schuchman E. H., Stramm L. E., Concaugh E. A., Haskins M. E., Aguirre G. D., Patterson D. F., Desnick R. J., Gilboa E. Restoration of normal lysosomal function in mucopolysaccharidosis type VII cells by retroviral vector-mediated gene transfer. Proc Natl Acad Sci U S A. 1990 Apr;87(8):2877–2881. doi: 10.1073/pnas.87.8.2877. [DOI] [PMC free article] [PubMed] [Google Scholar]