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- Bolshakova, Anastayia, et al.
(författare)
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EGF-induced dynamics of NF-kappa B and F-actin in A431 cells spread on fibronectin
- 2015
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Ingår i: Histochemistry and Cell Biology. - : Springer Verlag (Germany). - 0948-6143 .- 1432-119X. ; 144:3, s. 223-235
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Tidskriftsartikel (refereegranskat)abstract
- To evaluate the role of actin cytoskeleton in the regulation of NF-kappa B transcription factor, we analyzed its involvement in the intracellular transport and nuclear translocation of the NF-kappa B RelA/p65 subunit in A431 epithelial cells stimulated with fibronectin and EGF. Live cell imaging and confocal microscopy showed that EGF activated the movement of RelA/p65 in the cytoplasm. Upon cell adhesion to fibronectin, RelA/p65 concentrated onto stress fibers, and EGF stimulated its subsequent allocation to membrane ruffles, newly organized stress fibers, and discrete cytoplasmic actin-rich patches. These patches also contained alpha-actinin-1 and alpha-actinin-4, vinculin, paxillin, alpha-tubulin, and PI3-kinase. Cytochalasin D treatment resulted in RelA/p65 redistribution to actin-containing aggregates, with the number of cells with RelA/p65-containing clusters in the cytoplasm increasing under the effect of EGF. Furthermore, EGF proved to induce RelA/p65 accumulation in the nucleus after cell pretreatment with actin-stabilizing and actin-destabilizing agents, which was accompanied by changes in its DNA-binding activity after either EGF stimulation or cytochalasin D treatment. Thus, EGF treatment of A431 cells results in simultaneous nuclear RelA/p65 translocation and cytoplasmic redistribution, with part of RelA/p65 pool forming a very tight association with actin-rich structures. Apparently, nuclear transport is independent on drug stabilization or destabilization of the actin.
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