Intermediate filament protein synemin is present in human reactive and malignant astrocytes and associates with ruffled membranes in astrocytoma cells.
Synemin, a very unique type VI intermediate filament (IF) protein, exhibits alternative splice variants termed alpha and beta. Unlike other IF proteins, synemin binds to actin-associated proteins, including alpha-actinin, vinculin, and alpha-dystrobrevin. Our previous work has demonstrated the presence of synemin in differentiating astrocytes. In this study, we have examined the presence of synemin in human astrocytes under pathological conditions, using rabbit antibodies raised against the C-terminal domain of human synemin produced in bacteria. Western blotting shows that astrocytic tumors contain greater amounts of alpha-synemin than do normal brain tissues. These tumors also contain beta-synemin, which is not detectable in normal brain. Immunohistochemistry demonstrates that, while synemin is present in normal adult brain only in vascular smooth muscle cells, it is newly synthesized by reactive and neoplastic astrocytes. Alpha- and beta-Synemins have also been detected by Western blotting and polymerase chain reaction in several human glioblastoma cell lines. In these cell lines, surprisingly, synemin is associated with ruffled membranes in addition to being distributed along the IF network. In ruffled membranes, synemin was found to co-localize with alpha-actinin. This unusual cellular localization for an IF protein is maintained after nocodazole-induced perinuclear coiling of the vimentin IF network. In addition, immunoprecipitation experiments demonstrate that synemin forms a complex with alpha-actinin in glioblastoma cells. Taken together with synemin localization within ruffled membranes, this finding suggests that synemin plays a role in motility of glioblastoma cells.
Jing
Runfeng
R
Department of Cellular Biology and Anatomy and Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA.
Pizzolato
Gianpaolo
G
Robson
Richard M
RM
Gabbiani
Giulio
G
Skalli
Omar
O
D000185:Actinin / Q000378:metabolism; D000912:Antibodies, Neoplasm / Q000096:biosynthesis / Q000302:isolation & purification; D001253:Astrocytes / Q000378:metabolism*; D001254:Astrocytoma / Q000378:metabolism*; D015153:Blotting, Western; D001923:Brain Chemistry; D001932:Brain Neoplasms / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D009124:Muscle Proteins / Q000276:immunology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014746:Vimentin / Q000378:metabolism
D000912:Antibodies, Neoplasm; D007381:Intermediate Filament Proteins; D009124:Muscle Proteins; D014746:Vimentin; C430456:desmuslin; D000185:Actinin