The p65/RelA subunit of NF-kappaB interacts with actin-containing structures.
Nuclear factor-kappa B (NF-kappaB) is a universal transcription factor that participates in induction of a wide variety of cellular genes. In nonstimulated cells, NF-kappaB is sequestered in the cytoplasm. However, little is known about where NF-kappaB is located. We have studied the effect of inducing a reorganization of the actin filament system on NF-kappaB distribution, using normal and E1A+cHa-ras-transformed rat fibroblasts. This paper demonstrates that the p65/RelA subunit of NF-kappaB interacts with actin-containing structures. Immunofluorescence reveals that p65 is concentrated in focal contacts and along stress fibers in normal fibroblasts. Restoration of actin stress fibers in transformants spread on fibronectin is followed by reallocation of p65 to focal contacts and stress fibers, as in normal cells. The p65 is accumulated at the edge of leading lamellae in transformants spread on laminin and is redistributed to cell-to-cell contacts after a prolonged cultivation. Treatment of cells with Cytochalasin D leads to redistribution of p65 into the actin-containing aggregates. Affinity chromatography on matrix-bound F-actin confirms that p65 can bind to filamentous actin. Taken together, these data indicate that distribution of p65 in the cytoplasm depends on the state of the actin cytoskeleton and suggest an additional, yet unknown, function of the NF-kappaB in the cytoplasm.
Are
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AF
Department of Cell Cultures, Institute of Cytology Russian Academy of Sciences, Sankt-Petersburg, 194064, Russia.
Galkin
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VE
Pospelova
T V
TV
Pinaev
G P
GP
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D015638:Cytochalasin D / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D005353:Fibronectins / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D007797:Laminin / Q000502:physiology; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000378:metabolism*; D051381:Rats; D051996:Transcription Factor RelA
D000199:Actins; D005353:Fibronectins; D007797:Laminin; D016328:NF-kappa B; D051996:Transcription Factor RelA; D015638:Cytochalasin D