Skip to Content
MilliporeSigma
  • Raf-1/bcl-2 phosphorylation: a step from microtubule damage to cell death.

Raf-1/bcl-2 phosphorylation: a step from microtubule damage to cell death.

Cancer research (1997-01-01)
M V Blagosklonny, P Giannakakou, W S el-Deiry, D G Kingston, P I Higgs, L Neckers, T Fojo
ABSTRACT

Recent studies have shown that paclitaxel leads to activation of Raf-1 kinase and have suggested that this activation is essential for bcl-2 phosphorylation and apoptosis. In the present study, we demonstrate that, in addition to paclitaxel, other agents that interact with tubulin and microtubules also induce Raf-1/bcl-2 phosphorylation, whereas DNA-damaging drugs, antimetabolites, and alkylating agents do not. Activation of Raf-1 kinase by paclitaxel is linked to tubulin polymerization; the effect is blunted in paclitaxel-resistant cells, the tubulin of which does not polymerize following the addition of paclitaxel. In contrast, vincristine and vinblastine, drugs to which the paclitaxel-resistant cells retain sensitivity were able to bring about Raf-1 phosphorylation. The requirement for disruption of microtubules in this signaling cascade was strengthened further using paclitaxel analogues by demonstrating a correlation between tubulin polymerization, Raf-1/bcl-2 phosphorylation, and cytotoxicity. Inhibition of RNA or protein synthesis prevents Raf-1 activation and bcl-2 phosphorylation, suggesting that an intermediate protein(s) acts upstream of Raf-1 in this microtubule damage-activating pathway. A model is proposed that envisions a pathway of Raf-1 activation and bcl-2 phosphorylation following disruption of microtubular architecture, serving a role similar to p53 induction following DNA damage.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Vinblastine sulfate salt, ≥97% (HPLC)
Sigma-Aldrich
Vincristine sulfate salt, 95.0-105.0% (HPLC), powder or crystals