Subcellular localization of spastin: implications for the pathogenesis of hereditary spastic paraplegia.
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.
Svenson
Ingrid K
IK
Department of Molecular Genetics and Microbiology, Duke University Medical Center, P.O. Box 3175, Durham, NC 27710, USA.
Kloos
Mark T
MT
Jacon
Amy
A
Gallione
Carol
C
Horton
April C
AC
Pericak-Vance
Margaret A
MA
Ehlers
Michael D
MD
Marchuk
Douglas A
DA
D000251:Adenosine Triphosphatases / Q000235:genetics*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D006801:Humans; D008870:Microtubules / Q000473:pathology; D010264:Paraplegia / Q000235:genetics* / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000032:analysis; D000074181:Spastin; D014162:Transfection
D011993:Recombinant Fusion Proteins; D000251:Adenosine Triphosphatases; D000074181:Spastin; C400283:SPAST protein, human