EGF-induced dynamics of NF-kappa B and F-actin in A431 cells spread on fibronectin
2015 (English)In: Histochemistry and Cell Biology, ISSN 0948-6143, E-ISSN 1432-119X, Vol. 144, no 3, 223-235 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
Springer Verlag (Germany) , 2015. Vol. 144, no 3, 223-235 p.
RelA/p65; Actin cytoskeleton; Fibronectin; EGF; Cytochalasin D; Jasplakinolide
IdentifiersURN: urn:nbn:se:liu:diva-121098DOI: 10.1007/s00418-015-1331-5ISI: 000359650000003PubMedID: 25990946OAI: oai:DiVA.org:liu-121098DiVA: diva2:851859
Funding Agencies|Swedish Institute [879/2009]; wedish Research Council [2010-3045]; Faculty of Health Science, Linkoping University; European Science Foundation; Molecular and Cellular Biology Program of Russian Academy of Sciences; Russian Foundation for Basic Research [13-04-00497]2015-09-072015-09-072016-04-12