IN-VIVO CONTROL OF NF-KAPPA-B ACTIVATION BY I-KAPPA-B-ALPHA

The transcription factor NF-chiB is stored in the cytoplasm in complex es with the inhibitor protein IchiBalpha. It has been shown in vitro t hat dissociation of IchiBalpha from these complexes results in active NF-chiB. In this report we show that lipopolysaccharide (LPS)-induced activation of B or pre-B cells results in loss of IchiBalpha from NF-c hiB complexes in vivo. Many liberated NF-chiB dimers reached the nucle us, where increased c-rel, p65 and p50 were detected by immunoblotting and by DNA binding assays. Some liberated dimers were retained in the cytoplasm, however, through binding to newly synthesized IchiBalpha, a finding which strongly suggests (i) that the LPS-induced signal caus es dissociation of complexes rather than preventing their association and (ii) that dissociation results from modification of IchiBalpha and not of c-rel or p65. No effect of LPS treatment was detected on p105 or p100, which also retain rel family members in the cytoplasm. Quite unexpectedly, we also found that in unstimulated cells there is a cons tant ongoing process of degradation and replacement of complexed IchiB alpha. We propose that this turnover results in the low level of activ e NF-chiB presumably necessary even in the unstimulated cell, and that the high rate of synthesis of IchiBalpha provides the ability to turn off NF-chiB activity rapidly as soon as the activating signal ceases.