CARD-8 protein, a new CARD family member that regulates caspase-1 activation and apoptosis

Caspase-associated recruitment domains (CARD) are protein-protein interaction modules found extensively in proteins that play important roles in Apoptosis, NFkappaB activation, and cytokine regulation. In this study we identified a novel human protein, CARD-8, which contains a C-terminal CARD domain with high similarity to the CARD domain of Caspase-1/ICE. We demonstrate that CARD-8 interacts physically with Caspase-1 and negatively regulates caspase-1-dependent IL-1beta generation in the THP-1 monocytic cell line. CARD-8 binds also to ICEBERG and pseudo-ICE, two other recently identified proteins, which bind to the CARD domain of Caspase-1 and negatively regulate its activity. Reverse transcriptase-PCR analysis revealed that CARD-8 is expressed mainly in monocytes, placenta, lymph nodes, and spleen. This pattern of expression is consistent with Caspase-1 expression in the same cells and tissues. CARD-8 was also found to negatively regulate NF-kappaB activation by TNF-alpha stimulation and by ectopically expressed RICK, suggesting that this protein may control cell survival. Consistent with these results, stable expression of CARD-8 in U937 or THP-1 cells sensitizes the cells to differentiation-induced Apoptosis. Overexpression of CARD-8 can also induce Apoptosis in transfected cells. The results suggest that CARD-8 represents a new signaling molecule involved in the regulation of Caspase-1 and NF-kappaB activation.