Binding of kinesin to stress fibers in fibroblasts under condition of microtubule depolymerization.
The localization of kinesin in EBTr (bovine embryonic trachea fibroblast) cells was studied by indirect immunofluorescence microscopy using an affinity-purified antibody against bovine adrenal kinesin. It has already been shown that in interphase cells a part of kinesin is located on microtubules and the rest diffusely distributed throughout the cytoplasm [Murofushi et al., 1988]. When microtubules were depolymerized with cold or colchicine treatment, antikinesin antibody-stained fibrous components distinct from microtubules. These fibrous structures were considered to be stress fibers because they were stained with rhodamine-phalloidin and because the fibrous staining with antikinesin antibody was completely lost by treating the cells with cytochalasin D along with colchicine. When cold-treated cells in which a major part of kinesin had been localized on stress fibers were incubated at 37 degrees C, kinesin reappeared on reconstituted microtubules. These observations strongly suggest that kinesin has affinity not only to microtubules but also to stress fibers in culture cells.
Okuhara
K
K
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
Murofushi
H
H
Sakai
H
H
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002417:Cattle; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D003080:Cold Temperature; D003593:Cytoplasm / Q000378:metabolism; D005347:Fibroblasts / Q000032:analysis / Q000378:metabolism*; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000378:metabolism*; D016547:Kinesins; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D009419:Nerve Tissue Proteins; D016547:Kinesins; D003078:Colchicine