The Drosophila Homologue of the Hereditary Spastic Paraplegia Protein, Spastin, Severs and Disassembles Microtubules

Curr Biol. 2005 Apr 12;15(7):650-5. doi: 10.1016/j.cub.2005.02.029.

Abstract

Hereditary spastic paraplegias (HSPs), a group of neurodegenerative disorders characterized by lower-extremity spasticity and weakness, are most commonly caused by mutations in the spastin gene, which encodes a AAA+ ATPase related to the microtubule-severing protein katanin. A Drosophila homolog of spastin (D-spastin) was identified recently, and D-spastin RNAi-treated or genetic null flies show neurological defects, and protein overexpression decreases the density of cellular microtubules. Elucidating spastin's function and disease mechanism will require a more detailed understanding of its structure and biochemical mechanism. Here, we have investigated the effects of D-spastin, individual D-spastin domains, and D-spastin proteins bearing disease mutations on microtubules in cellular and in vitro assays. We show that D-spastin, like katanin, displays ATPase activity and uses energy from ATP hydrolysis to sever and disassemble microtubules; disease mutations abolish or partially interfere with these activities.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / genetics*
  • Adenosine Triphosphatases / metabolism*
  • Animals
  • Cell Line
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / genetics*
  • Escherichia coli
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Immunoblotting
  • Katanin
  • Microtubules / metabolism*
  • Mutation / genetics
  • Protein Structure, Tertiary
  • RNA Interference
  • Sequence Analysis, DNA
  • Spastic Paraplegia, Hereditary / genetics*

Substances

  • DNA, Complementary
  • Drosophila Proteins
  • Kat60 protein, Drosophila
  • Green Fluorescent Proteins
  • Adenosine Triphosphatases
  • spas protein, Drosophila
  • Katanin