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. 1989 Nov;8(11):3295–3302. doi: 10.1002/j.1460-2075.1989.tb08490.x

Myelin-deficient rat: a point mutation in exon III (A----C, Thr75----Pro) of the myelin proteolipid protein causes dysmyelination and oligodendrocyte death.

D Boison 1, W Stoffel 1
PMCID: PMC401462  PMID: 2479544

Abstract

The expression of the proteolipid protein (PLP) gene of the myelin deficient (md) and normal rat was studied during the myelination period. The sizes of the PLP transcripts (1.6 and 3.2 kb) in the md and normal rat were identical although the md PLP messenger RNA level was extremely reduced as shown by in situ hybridization and Northern blot hybridization analysis. The structure of the md proteolipid protein gene was analyzed on the cDNA and genomic level. The molecular basis of the myelin deficiency phenotype has been elucidated: a point mutation in exon III (A----C transversion) verified by cDNA and genomic DNA sequencing causes a mutation of Thr75 to Pro and creates an additional AvaII restriction site in exon III of the md rat. The threonine to proline mutation located within the second transmembranal alpha-helix might induce a conformational change and thereby prohibit the integration of PLP into the membrane with the clinical manifestation of dysmyelination leading to premature death within 3-6 weeks.

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Selected References

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

  1. Blobel G. Control of intracellular protein traffic. Methods Enzymol. 1983;96:663–682. doi: 10.1016/s0076-6879(83)96056-1. [DOI] [PubMed] [Google Scholar]
  2. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  3. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Csiza C. K., de Lahunta A. Myelin deficiency (md): a neurologic mutant in the Wistar rat. Am J Pathol. 1979 Apr;95(1):215–224. [PMC free article] [PubMed] [Google Scholar]
  6. Dentinger M. P., Barron K. D., Csiza C. K. Glial and axonal development in optic nerve of myelin deficient rat mutant. Brain Res. 1985 Oct 7;344(2):255–266. doi: 10.1016/0006-8993(85)90803-0. [DOI] [PubMed] [Google Scholar]
  7. Dentinger M. P., Barron K. D., Csiza C. K. Ultrastructure of the central nervous system in a myelin deficient rat. J Neurocytol. 1982 Aug;11(4):671–691. doi: 10.1007/BF01262431. [DOI] [PubMed] [Google Scholar]
  8. Diehl H. J., Schaich M., Budzinski R. M., Stoffel W. Individual exons encode the integral membrane domains of human myelin proteolipid protein. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9807–9811. doi: 10.1073/pnas.83.24.9807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duncan I. D., Hammang J. P., Trapp B. D. Abnormal compact myelin in the myelin-deficient rat: absence of proteolipid protein correlates with a defect in the intraperiod line. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6287–6291. doi: 10.1073/pnas.84.17.6287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  12. Hudson L. D., Berndt J. A., Puckett C., Kozak C. A., Lazzarini R. A. Aberrant splicing of proteolipid protein mRNA in the dysmyelinating jimpy mutant mouse. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1454–1458. doi: 10.1073/pnas.84.5.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knapp P. E., Skoff R. P., Redstone D. W. Oligodendroglial cell death in jimpy mice: an explanation for the myelin deficit. J Neurosci. 1986 Oct;6(10):2813–2822. doi: 10.1523/JNEUROSCI.06-10-02813.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kumar S., Gordon M. N., Espinosa de los Monteros M. A., de Vellis J. Developmental expression of neural cell type-specific mRNA markers in the myelin-deficient mutant rat brain: inhibition of oligodendrocyte differentiation. J Neurosci Res. 1988 Oct-Dec;21(2-4):268–274. doi: 10.1002/jnr.490210219. [DOI] [PubMed] [Google Scholar]
  15. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  16. Morello D., Dautigny A., Pham-Dinh D., Jollès P. Myelin proteolipid protein (PLP and DM-20) transcripts are deleted in jimpy mutant mice. EMBO J. 1986 Dec 20;5(13):3489–3493. doi: 10.1002/j.1460-2075.1986.tb04674.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nave K. A., Lai C., Bloom F. E., Milner R. J. Jimpy mutant mouse: a 74-base deletion in the mRNA for myelin proteolipid protein and evidence for a primary defect in RNA splicing. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9264–9268. doi: 10.1073/pnas.83.23.9264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  19. SIDMAN R. L., DICKIE M. M., APPEL S. H. MUTANT MICE (QUAKING AND JIMPY) WITH DEFICIENT MYELINATION IN THE CENTRAL NERVOUS SYSTEM. Science. 1964 Apr 17;144(3616):309–311. doi: 10.1126/science.144.3616.309. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  22. Schaich M., Budzinski R. M., Stoffel W. Cloned proteolipid protein and myelin basic protein cDNA. Transcription of the two genes during myelination. Biol Chem Hoppe Seyler. 1986 Aug;367(8):825–834. doi: 10.1515/bchm3.1986.367.2.825. [DOI] [PubMed] [Google Scholar]
  23. Stoffel W., Hillen H., Giersiefen H. Structure and molecular arrangement of proteolipid protein of central nervous system myelin. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5012–5016. doi: 10.1073/pnas.81.16.5012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stoffel W., Hillen H., Schröder W., Deutzmann R. The primary structure of bovine brain myelin lipophilin (proteolipid apoprotein). Hoppe Seylers Z Physiol Chem. 1983 Oct;364(10):1455–1466. doi: 10.1515/bchm2.1983.364.2.1455. [DOI] [PubMed] [Google Scholar]
  25. Tourtellotte W. W., Verity A. N., Schmid P., Martinez S., Shapshak P. Covalent binding of formalin fixed paraffin embedded brain tissue sections to glass slides suitable for in situ hybridization. J Virol Methods. 1987 Feb;15(2):87–99. doi: 10.1016/0166-0934(87)90052-8. [DOI] [PubMed] [Google Scholar]
  26. Willard H. F., Riordan J. R. Assignment of the gene for myelin proteolipid protein to the X chromosome: implications for X-linked myelin disorders. Science. 1985 Nov 22;230(4728):940–942. doi: 10.1126/science.3840606. [DOI] [PubMed] [Google Scholar]
  27. Yanagisawa K., Duncan I. D., Hammang J. P., Quarles R. H. Myelin-deficient rat: analysis of myelin proteins. J Neurochem. 1986 Dec;47(6):1901–1907. doi: 10.1111/j.1471-4159.1986.tb13105.x. [DOI] [PubMed] [Google Scholar]

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