UVB LIGHT INDUCES NUCLEAR FACTOR KB (NFKB) ACTIVITY INDEPENDENTLY FROM CHROMOSOMAL DNA-DAMAGE IN CELL-FREE CYTOSOLIC EXTRACTS

It has been shown previously that ultraviolet (UV) light (290 - 320 nm ) activates keratinocytes to release proinflammatory cytokines includi ng interleukin (IL)-6. Because the 5' flanking region of the IL-6 gene contains a consensus NF kappa B binding sequence, the effect of UVB l ight on an NF kappa B-like binding activity was investigated in a huma n epidermoid carcinoma cell line (A431). Nuclear factor kappa B (NF ka ppa B) activation in the cytoplasm is known to be due to the dissociat ion of an inactive NF kappa B-inhibitor of nuclear factor kappa B (I k appa B) complex. Cytosolic extracts from cells harvested shortly after sublethal UVB irradiation showed a UVB dose-dependent increase of NF kappa B binding The activation was reduced by radical scavenging chemi cals, suggesting involvement of reactive oxygen intermediates. NF kapp a B activation has been shown previously to be triggered by DNA lesion s induced by UV light. To elucidate whether DNA damage is necessary an d sufficient to mediate NF kappa B activation crude, cytosolic protein extracts obtained from unirradiated cells were exposed to UVB light. This in vitro UVB treatment led to activation of an NF kappa B-like bi nding activity, suggesting an additional signaling pathway independent of chromosomal DNA damage or byproducts of DNA damage. The activation process was dependent on the presence of membranes. The data suggest at least an additional signaling pathway for the early UVB response, i ncluding a component of the pathway residing at the cell membrane.