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Neurogenetics

, Volume 6, Issue 3, pp 135–141 | Cite as

Subcellular localization of spastin: implications for the pathogenesis of hereditary spastic paraplegia

  • Ingrid K. Svenson
  • Mark T. Kloos
  • Amy Jacon
  • Carol Gallione
  • April C. Horton
  • Margaret A. Pericak-Vance
  • Michael D. Ehlers
  • Douglas A. MarchukEmail author
Original Article

Abstract

Hereditary spastic paraplegia (HSP) is a group of clinically and genetically heterogeneous diseases characterized by neuronal degeneration that is maximal at the distal ends of the longest axons of the central nervous system. The most common cause of autosomal dominant HSP is mutation of a novel gene encoding spastin, a protein whose function is still being elucidated. One clue concerning spastin function is its intracellular localization. Here, we describe a novel anti-spastin antiserum designed to a unique epitope contained within all splicing isoforms. The antiserum exhibits specific immunostaining of recombinant spastin in intact, fixed cells. Using this reagent, we find that endogenous spastin is located at the centrosome in a variety of cell types at all points in the cell cycle. This localization is resistant to microtubule disruption, suggesting that spastin may be an integral centrosomal protein. In addition to the centrosome, spastin also localizes at discrete focal regions along the axons of primary cultured neurons. These data lend additional support to the emerging hypothesis that spastin plays a role in microtubule dynamics, with a crucial role in microtubule organization.

Keywords

Nocodazole Microtubule Dynamic Hereditary Spastic Paraplegia Microtubule Network Microtubule Depolymerization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported by grant P01 NS26630 to M.P.V. I.K.S. was the recipient of a Ruth K. Broad Biomedical Research Foundation Fellowship Award.

References

  1. 1.
    Fink JK (1997) Advances in hereditary spastic paraplegia. Curr Opin Neurol 10:313–318PubMedCrossRefGoogle Scholar
  2. 2.
    Reid E (2003) Science in motion: common molecular pathological themes emerge in the hereditary spastic paraplegias. J Med Genet 40:81–86CrossRefPubMedGoogle Scholar
  3. 3.
    Hazan J, Fonknechten N, Mavel D, Paternotte C, Samson D, Artiguenave F, Davoine CS, Cruaud C, Durr A, Wincker P, Brottier P, Cattolico L, Barbe V, Burgunder JM, Prud’homme JF, Brice A, Fontaine B, Heilig B, Weissenbach J (1999) Spastin, a new AAA protein, is altered in the most frequent form of autosomal dominant spastic paraplegia. Nat Genet 23:296–303CrossRefPubMedGoogle Scholar
  4. 4.
    Patel S, Latterich M (1998) The AAA team: related ATPases with diverse functions. Trends Cell Biol 8:65–71CrossRefPubMedGoogle Scholar
  5. 5.
    Lupas AN, Martin J (2002) AAA proteins. Curr Opin Struct Biol 12:746–753CrossRefPubMedGoogle Scholar
  6. 6.
    Beetz C, Brodhun M, Moutzouris K, Kiehntopf M, Berndt A, Lehnert D, Deufel T, Bastmeyer M, Schickel J (2004) Identification of nuclear localisation sequences in spastin (SPG4) using a novel Tetra-GFP reporter system. Biochem Biophys Res Commun 318:1079–1084CrossRefPubMedGoogle Scholar
  7. 7.
    Ciccarelli FD, Proukakis C, Patel H, Cross H, Azam S, Patton MA, Bork P, Crosby AH (2003) The identification of a conserved domain in both spartin and spastin, mutated in hereditary spastic paraplegia. Genomics 81:437–441CrossRefPubMedGoogle Scholar
  8. 8.
    Errico A, Ballabio A, Rugarli EI (2002) Spastin, the protein mutated in autosomal dominant hereditary spastic paraplegia, is involved in microtubule dynamics. Hum Mol Genet 11:153–163CrossRefPubMedGoogle Scholar
  9. 9.
    McDermott CJ, Grierson AJ, Wood JD, Bingley M, Wharton SB, Bushby KM, Shaw PJ (2003) Hereditary spastic paraparesis: disrupted intracellular transport associated with spastin mutation. Ann Neurol 54:748–759CrossRefPubMedGoogle Scholar
  10. 10.
    Errico A, Claudiani P, D’Addio M, Rugarli EI (2004) Spastin interacts with the centrosomal protein NA14, and is enriched in the spindle pole, the midbody and the distal axon. Hum Mol Genet 13:2121–2132CrossRefPubMedGoogle Scholar
  11. 11.
    Reid E, Connell J, Edwards TL, Duley S, Brown SE, Sanderson CM (2004) The hereditary spastic paraplegia protein spastin interacts with the ESCRT-III complex-associated endosomal protein CHMP1B. Hum Mol GenetGoogle Scholar
  12. 12.
    Charvin D, Cifuentes-Diaz C, Fonknechten N, Joshi V, Hazan J, Melki J, Betuing S (2003) Mutations of SPG4 are responsible for a loss of function of spastin, an abundant neuronal protein localized in the nucleus. Hum Mol Genet 12:71–78CrossRefPubMedGoogle Scholar
  13. 13.
    Wharton SB, McDermott CJ, Grierson AJ, Wood JD, Gelsthorpe C, Ince PG, Shaw PJ (2003) The cellular and molecular pathology of the motor system in hereditary spastic paraparesis due to mutation of the spastin gene. J Neuropathol Exp Neurol 62:1166–1177PubMedGoogle Scholar
  14. 14.
    Svenson IK, Ashley-Koch AE, Gaskell PC, Riney TJ, Cumming WJ, Kingston HM, Hogan EL, Boustany RM, Vance JM, Nance MA, Pericak-Vance MA, Marchuk DA (2001) Identification and expression analysis of spastin gene mutations in hereditary spastic paraplegia. Am J Hum Genet 68:1077–1085CrossRefPubMedGoogle Scholar
  15. 15.
    Trinczek B, Brajenovic M, Ebneth A, Drewes G (2004) MARK4 is a novel microtubule-associated proteins/microtubule affinity-regulating kinase that binds to the cellular microtubule network and to centrosomes. J Biol Chem 279:5915–5923CrossRefPubMedGoogle Scholar
  16. 16.
    Reid E, Kloos M, Ashley-Koch A, Hughes L, Bevan S, Svenson IK, Graham FL, Gaskell PC, Dearlove A, Pericak-Vance MA, Rubinsztein DC, Marchuk DA (2002) A kinesin heavy chain (KIF5A) mutation in hereditary spastic paraplegia (SPG10). Am J Hum Genet 71:1189–1194CrossRefPubMedGoogle Scholar
  17. 17.
    Crosby AH, Proukakis C (2002) Is the transportation highway the right road for hereditary spastic paraplegia? Am J Hum Genet 71:1009–1016CrossRefPubMedGoogle Scholar
  18. 18.
    Frickey T, Lupas AN (2004) Phylogenetic analysis of AAA proteins. J Struct Biol 146:2–10CrossRefPubMedGoogle Scholar
  19. 19.
    Ahmad FJ, Yu W, McNally FJ, Baas PW (1999) An essential role for katanin in severing microtubules in the neuron. J Cell Biol 145:305–315CrossRefPubMedGoogle Scholar
  20. 20.
    Kimble M, Kuriyama R (1992) Functional components of microtubule-organizing centers. Int Rev Cytol 136:1–50PubMedCrossRefGoogle Scholar
  21. 21.
    McNally FJ, Vale RD (1993) Identification of katanin, an ATPase that severs and disassembles stable microtubules. Cell 75:419–429CrossRefPubMedGoogle Scholar
  22. 22.
    McNally FJ, Okawa K, Iwamatsu A, Vale RD (1996) Katanin, the microtubule-severing ATPase, is concentrated at centrosomes. J Cell Sci 109:561–567PubMedGoogle Scholar
  23. 23.
    Horton AC, Ehlers MD (2003) Dual modes of endoplasmic reticulum-to-Golgi transport in dendrites revealed by live-cell imaging. J Neurosci 23:6188–6199PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Ingrid K. Svenson
    • 1
  • Mark T. Kloos
    • 1
  • Amy Jacon
    • 1
  • Carol Gallione
    • 1
  • April C. Horton
    • 2
  • Margaret A. Pericak-Vance
    • 3
  • Michael D. Ehlers
    • 2
  • Douglas A. Marchuk
    • 1
    Email author
  1. 1.Department of Molecular Genetics and MicrobiologyDuke University Medical CenterDurhamUSA
  2. 2.Department of NeurobiologyDuke University Medical CenterDurhamUSA
  3. 3.Department of Medicine, Center for Human GeneticsDuke University Medical CenterDurhamUSA

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