Extracellular ATP Activates Transcription Factor NF-κB through the P2Z Purinoreceptor by Selectively Targeting NF-κB p65 (RelA)

Cells of the macrophage lineage express a peculiar surface receptor for extracellular ATP, designated P2Z/P2X₇ purinergic receptor, that induces pore formation and collapse of the plasma membrane potential. Although the function of the P2Z receptor is largely unknown, accumulating evidence implicates its role in cell signaling and immune reactions. Here, we investigated the effect of P2Z receptor ligation on the activation of NF-κB, a transcription factor controlling cytokine expression and apoptosis. Exposure of microglial cells to ATP but not other nucleotides resulted in potent NF-κB activation. This effect was specifically mediated by the P2Z receptor, because selective receptor antagonists prevented NF-κB activation. NF-κB activation required reactive oxygen intermediates and proteases of the caspase family, because it was abolished by antioxidants and specific protease inhibitors. The subunit composition of the ATP-induced NF- κB–DNA complex was rather unusual. Whereas exposure to LPS-induced prototypical NF-κB p50 homo- and p65 (RelA)/p50 heterodimers, ATP stimulation resulted in the sole appearance of a p65 homodimer. This is the first demonstration that a certain stimulus activates a particular NF-κB subunit. Because different NF-κB complexes exhibit distinct transcriptional and DNA-binding activities, ATP may control the expression of a subset of NF-κB target genes distinct from those activated by classical proinflammatory mediators.