Erythroid gene suppression by NF-κB

NF-kappaB/Rel transcription factors play essential roles to mediate the immune response and apoptosis, and they have also been implicated in cellular differentiation such as erythropoiesis. To elucidate the possible role(s) of NF-kappaB in erythroid gene regulation and erythropoiesis, we have carried out transient transfection studies of the human embryonic/ fetal erythroid cell line K562 and mouse adult erythroid MEL cells. It is shown that tumor necrosis factor-alpha represses the transcription activity directed by either alpha or zeta globin promoter in a dose-dependent manner. Furthermore, different NF-kappaB family members could effectively repress the transfected alpha-like globin promoters in K562 as well as in MEL cells. The involvement of NF-kappaB pathway is supported by the ability of a NF-kappaB-specific, dominant negative mutant to block the tumor necrosis factor-alpha or p65-mediated suppression of the alpha-like globin promoter activities. The suppression appears to be mediated through cis-linked HS-40 enhancer. Finally, stably transfected K562 cells overexpressing p65 contain reduced amounts of the p45/ NF-E2 RNA and functional NF-E2 proteins. Our studies have identified a new set of targets of NF-kappaB. We suggest that the relatively high activity of the NF-kappaB pathway in early erythroid progenitors is involved in the suppression of erythroid-specific genes. Later in differentiation, together with other changes, the decline of the amounts of the NF-kappaB family of factors leads to derepression and consequent increase of NF-E2, which in turn would activate a subset of erythroid-specific genes.