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The Raf-1 kinase associates with vimentin kinases and regulates the structure of vimentin filaments

By Petra Janosch, Arnd Kieser, Manfred Eulitz, Josip Lovric, Guido Sauer, Manuela Reichert, Fotini Gounari, Dirk Büscher, Manuela Baccarini, Harald Mischak and Walter Kolch


ABSTRACT Using immobilized GST-Raf-1 as bait, we have isolated the intermediate filament protein vimentin as a Raf-1-associated protein. Vimentin coimmunoprecipitated and colocalized with Raf-1 in fibroblasts. Vimentin was not a Raf-1 substrate, but was phosphorylated by Raf-1-associated vimentin kinases. We provide evidence for at least two Raf-1associated vimentin kinases and identified one as casein kinase 2. They are regulated by Raf-1, since the activation status of Raf-1 correlated with the phosphorylation of vimentin. Vimentin phosphorylation by Raf-1 preparations interfered with its polymerization in vitro. A subset of tryptic vimentin phosphopeptides induced by Raf-1 in vitro matched the vimentin phosphopeptides isolated from v-raftransfecte

Year: 2000
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