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. 1996 Aug 1;184(2):609–618. doi: 10.1084/jem.184.2.609

Selective loss of sarcolemmal nitric oxide synthase in Becker muscular dystrophy

PMCID: PMC2192729  PMID: 8760814

Abstract

Becker muscular dystrophy is an X-linked disease due to mutations of the dystrophin gene. We now show that neuronal-type nitric oxide synthase (nNOS), an identified enzyme in the dystrophin complex, is uniquely absent from skeletal muscle plasma membrane in many human Becker patients and in mouse models of dystrophinopathy. An NH2- terminal domain of nNOS directly interacts with alpha 1-syntrophin but not with other proteins in the dystrophin complex analyzed. However, nNOS does not associate with alpha 1-syntrophin on the sarcolemma in transgenic mdx mice expressing truncated dystrophin proteins. This suggests a ternary interaction of nNOS, alpha 1-syntrophin, and the central domain of dystrophin in vivo, a conclusion supported by developmental studies in muscle. These data indicate that proper assembly of the dystrophin complex is dependent upon the structure of the central rodlike domain and have implications for the design of dystrophin-containing vectors for gene therapy.

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Selected References

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  1. Adams M. E., Butler M. H., Dwyer T. M., Peters M. F., Murnane A. A., Froehner S. C. Two forms of mouse syntrophin, a 58 kd dystrophin-associated protein, differ in primary structure and tissue distribution. Neuron. 1993 Sep;11(3):531–540. doi: 10.1016/0896-6273(93)90157-m. [DOI] [PubMed] [Google Scholar]
  2. Adams M. E., Dwyer T. M., Dowler L. L., White R. A., Froehner S. C. Mouse alpha 1- and beta 2-syntrophin gene structure, chromosome localization, and homology with a discs large domain. J Biol Chem. 1995 Oct 27;270(43):25859–25865. doi: 10.1074/jbc.270.43.25859. [DOI] [PubMed] [Google Scholar]
  3. Ahn A. H., Kunkel L. M. Syntrophin binds to an alternatively spliced exon of dystrophin. J Cell Biol. 1995 Feb;128(3):363–371. doi: 10.1083/jcb.128.3.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arahata K., Hoffman E. P., Kunkel L. M., Ishiura S., Tsukahara T., Ishihara T., Sunohara N., Nonaka I., Ozawa E., Sugita H. Dystrophin diagnosis: comparison of dystrophin abnormalities by immunofluorescence and immunoblot analyses. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7154–7158. doi: 10.1073/pnas.86.18.7154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beggs A. H., Hoffman E. P., Snyder J. R., Arahata K., Specht L., Shapiro F., Angelini C., Sugita H., Kunkel L. M. Exploring the molecular basis for variability among patients with Becker muscular dystrophy: dystrophin gene and protein studies. Am J Hum Genet. 1991 Jul;49(1):54–67. [PMC free article] [PubMed] [Google Scholar]
  6. Bredt D. S., Hwang P. M., Snyder S. H. Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature. 1990 Oct 25;347(6295):768–770. doi: 10.1038/347768a0. [DOI] [PubMed] [Google Scholar]
  7. Brenman J. E., Chao D. S., Gee S. H., McGee A. W., Craven S. E., Santillano D. R., Wu Z., Huang F., Xia H., Peters M. F. Interaction of nitric oxide synthase with the postsynaptic density protein PSD-95 and alpha1-syntrophin mediated by PDZ domains. Cell. 1996 Mar 8;84(5):757–767. doi: 10.1016/s0092-8674(00)81053-3. [DOI] [PubMed] [Google Scholar]
  8. Brenman J. E., Chao D. S., Xia H., Aldape K., Bredt D. S. Nitric oxide synthase complexed with dystrophin and absent from skeletal muscle sarcolemma in Duchenne muscular dystrophy. Cell. 1995 Sep 8;82(5):743–752. doi: 10.1016/0092-8674(95)90471-9. [DOI] [PubMed] [Google Scholar]
  9. Bönnemann C. G., Modi R., Noguchi S., Mizuno Y., Yoshida M., Gussoni E., McNally E. M., Duggan D. J., Angelini C., Hoffman E. P. Beta-sarcoglycan (A3b) mutations cause autosomal recessive muscular dystrophy with loss of the sarcoglycan complex. Nat Genet. 1995 Nov;11(3):266–273. doi: 10.1038/ng1195-266. [DOI] [PubMed] [Google Scholar]
  10. Campanelli J. T., Roberds S. L., Campbell K. P., Scheller R. H. A role for dystrophin-associated glycoproteins and utrophin in agrin-induced AChR clustering. Cell. 1994 Jun 3;77(5):663–674. doi: 10.1016/0092-8674(94)90051-5. [DOI] [PubMed] [Google Scholar]
  11. Cox G. A., Cole N. M., Matsumura K., Phelps S. F., Hauschka S. D., Campbell K. P., Faulkner J. A., Chamberlain J. S. Overexpression of dystrophin in transgenic mdx mice eliminates dystrophic symptoms without toxicity. Nature. 1993 Aug 19;364(6439):725–729. doi: 10.1038/364725a0. [DOI] [PubMed] [Google Scholar]
  12. Cox G. A., Sunada Y., Campbell K. P., Chamberlain J. S. Dp71 can restore the dystrophin-associated glycoprotein complex in muscle but fails to prevent dystrophy. Nat Genet. 1994 Dec;8(4):333–339. doi: 10.1038/ng1294-333. [DOI] [PubMed] [Google Scholar]
  13. Ervasti J. M., Campbell K. P. Dystrophin and the membrane skeleton. Curr Opin Cell Biol. 1993 Feb;5(1):82–87. doi: 10.1016/s0955-0674(05)80012-2. [DOI] [PubMed] [Google Scholar]
  14. Ervasti J. M., Kahl S. D., Campbell K. P. Purification of dystrophin from skeletal muscle. J Biol Chem. 1991 May 15;266(14):9161–9165. [PubMed] [Google Scholar]
  15. Ervasti J. M., Ohlendieck K., Kahl S. D., Gaver M. G., Campbell K. P. Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle. Nature. 1990 May 24;345(6273):315–319. doi: 10.1038/345315a0. [DOI] [PubMed] [Google Scholar]
  16. Gee S. H., Montanaro F., Lindenbaum M. H., Carbonetto S. Dystroglycan-alpha, a dystrophin-associated glycoprotein, is a functional agrin receptor. Cell. 1994 Jun 3;77(5):675–686. doi: 10.1016/0092-8674(94)90052-3. [DOI] [PubMed] [Google Scholar]
  17. Greenberg D. S., Sunada Y., Campbell K. P., Yaffe D., Nudel U. Exogenous Dp71 restores the levels of dystrophin associated proteins but does not alleviate muscle damage in mdx mice. Nat Genet. 1994 Dec;8(4):340–344. doi: 10.1038/ng1294-340. [DOI] [PubMed] [Google Scholar]
  18. Grozdanovic Z., Nakos G., Dahrmann G., Mayer B., Gossrau R. Species-independent expression of nitric oxide synthase in the sarcolemma region of visceral and somatic striated muscle fibers. Cell Tissue Res. 1995 Sep;281(3):493–499. doi: 10.1007/BF00417866. [DOI] [PubMed] [Google Scholar]
  19. Hoffman E. P., Fischbeck K. H., Brown R. H., Johnson M., Medori R., Loike J. D., Harris J. B., Waterston R., Brooke M., Specht L. Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne's or Becker's muscular dystrophy. N Engl J Med. 1988 May 26;318(21):1363–1368. doi: 10.1056/NEJM198805263182104. [DOI] [PubMed] [Google Scholar]
  20. Hoffman E. P. Genotype/phenotype correlations in Duchenne/Becker dystrophy. Mol Cell Biol Hum Dis Ser. 1993;3:12–36. doi: 10.1007/978-94-011-1528-5_2. [DOI] [PubMed] [Google Scholar]
  21. Hoffman E. P., Kunkel L. M., Angelini C., Clarke A., Johnson M., Harris J. B. Improved diagnosis of Becker muscular dystrophy by dystrophin testing. Neurology. 1989 Aug;39(8):1011–1017. doi: 10.1212/wnl.39.8.1011. [DOI] [PubMed] [Google Scholar]
  22. Hoffman E. P., Morgan J. E., Watkins S. C., Partridge T. A. Somatic reversion/suppression of the mouse mdx phenotype in vivo. J Neurol Sci. 1990 Oct;99(1):9–25. doi: 10.1016/0022-510x(90)90195-s. [DOI] [PubMed] [Google Scholar]
  23. Ibraghimov-Beskrovnaya O., Ervasti J. M., Leveille C. J., Slaughter C. A., Sernett S. W., Campbell K. P. Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix. Nature. 1992 Feb 20;355(6362):696–702. doi: 10.1038/355696a0. [DOI] [PubMed] [Google Scholar]
  24. Inglese J., Koch W. J., Touhara K., Lefkowitz R. J. G beta gamma interactions with PH domains and Ras-MAPK signaling pathways. Trends Biochem Sci. 1995 Apr;20(4):151–156. doi: 10.1016/s0968-0004(00)88992-6. [DOI] [PubMed] [Google Scholar]
  25. Kobzik L., Reid M. B., Bredt D. S., Stamler J. S. Nitric oxide in skeletal muscle. Nature. 1994 Dec 8;372(6506):546–548. doi: 10.1038/372546a0. [DOI] [PubMed] [Google Scholar]
  26. Koenig M., Monaco A. P., Kunkel L. M. The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell. 1988 Apr 22;53(2):219–228. doi: 10.1016/0092-8674(88)90383-2. [DOI] [PubMed] [Google Scholar]
  27. Lee K. H., Baek M. Y., Moon K. Y., Song W. K., Chung C. H., Ha D. B., Kang M. S. Nitric oxide as a messenger molecule for myoblast fusion. J Biol Chem. 1994 May 20;269(20):14371–14374. [PubMed] [Google Scholar]
  28. Lim L. E., Duclos F., Broux O., Bourg N., Sunada Y., Allamand V., Meyer J., Richard I., Moomaw C., Slaughter C. Beta-sarcoglycan: characterization and role in limb-girdle muscular dystrophy linked to 4q12. Nat Genet. 1995 Nov;11(3):257–265. doi: 10.1038/ng1195-257. [DOI] [PubMed] [Google Scholar]
  29. Noguchi S., McNally E. M., Ben Othmane K., Hagiwara Y., Mizuno Y., Yoshida M., Yamamoto H., Bönnemann C. G., Gussoni E., Denton P. H. Mutations in the dystrophin-associated protein gamma-sarcoglycan in chromosome 13 muscular dystrophy. Science. 1995 Nov 3;270(5237):819–822. doi: 10.1126/science.270.5237.819. [DOI] [PubMed] [Google Scholar]
  30. Peters M. F., Kramarcy N. R., Sealock R., Froehner S. C. beta 2-Syntrophin: localization at the neuromuscular junction in skeletal muscle. Neuroreport. 1994 Aug 15;5(13):1577–1580. [PubMed] [Google Scholar]
  31. Phelps S. F., Hauser M. A., Cole N. M., Rafael J. A., Hinkle R. T., Faulkner J. A., Chamberlain J. S. Expression of full-length and truncated dystrophin mini-genes in transgenic mdx mice. Hum Mol Genet. 1995 Aug;4(8):1251–1258. doi: 10.1093/hmg/4.8.1251. [DOI] [PubMed] [Google Scholar]
  32. Rafael J. A., Sunada Y., Cole N. M., Campbell K. P., Faulkner J. A., Chamberlain J. S. Prevention of dystrophic pathology in mdx mice by a truncated dystrophin isoform. Hum Mol Genet. 1994 Oct;3(10):1725–1733. doi: 10.1093/hmg/3.10.1725. [DOI] [PubMed] [Google Scholar]
  33. Roberds S. L., Leturcq F., Allamand V., Piccolo F., Jeanpierre M., Anderson R. D., Lim L. E., Lee J. C., Tomé F. M., Romero N. B. Missense mutations in the adhalin gene linked to autosomal recessive muscular dystrophy. Cell. 1994 Aug 26;78(4):625–633. doi: 10.1016/0092-8674(94)90527-4. [DOI] [PubMed] [Google Scholar]
  34. Suzuki A., Yoshida M., Hayashi K., Mizuno Y., Hagiwara Y., Ozawa E. Molecular organization at the glycoprotein-complex-binding site of dystrophin. Three dystrophin-associated proteins bind directly to the carboxy-terminal portion of dystrophin. Eur J Biochem. 1994 Mar 1;220(2):283–292. doi: 10.1111/j.1432-1033.1994.tb18624.x. [DOI] [PubMed] [Google Scholar]
  35. Suzuki A., Yoshida M., Ozawa E. Mammalian alpha 1- and beta 1-syntrophin bind to the alternative splice-prone region of the dystrophin COOH terminus. J Cell Biol. 1995 Feb;128(3):373–381. doi: 10.1083/jcb.128.3.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wang T., Xie Z., Lu B. Nitric oxide mediates activity-dependent synaptic suppression at developing neuromuscular synapses. Nature. 1995 Mar 16;374(6519):262–266. doi: 10.1038/374262a0. [DOI] [PubMed] [Google Scholar]
  37. Way M., Pope B., Cross R. A., Kendrick-Jones J., Weeds A. G. Expression of the N-terminal domain of dystrophin in E. coli and demonstration of binding to F-actin. FEBS Lett. 1992 Apr 27;301(3):243–245. doi: 10.1016/0014-5793(92)80249-g. [DOI] [PubMed] [Google Scholar]
  38. Wells D. J., Wells K. E., Asante E. A., Turner G., Sunada Y., Campbell K. P., Walsh F. S., Dickson G. Expression of human full-length and minidystrophin in transgenic mdx mice: implications for gene therapy of Duchenne muscular dystrophy. Hum Mol Genet. 1995 Aug;4(8):1245–1250. doi: 10.1093/hmg/4.8.1245. [DOI] [PubMed] [Google Scholar]
  39. Yang B., Ibraghimov-Beskrovnaya O., Moomaw C. R., Slaughter C. A., Campbell K. P. Heterogeneity of the 59-kDa dystrophin-associated protein revealed by cDNA cloning and expression. J Biol Chem. 1994 Feb 25;269(8):6040–6044. [PubMed] [Google Scholar]
  40. Yang B., Jung D., Rafael J. A., Chamberlain J. S., Campbell K. P. Identification of alpha-syntrophin binding to syntrophin triplet, dystrophin, and utrophin. J Biol Chem. 1995 Mar 10;270(10):4975–4978. doi: 10.1074/jbc.270.10.4975. [DOI] [PubMed] [Google Scholar]

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