In the present study, we describe the function of a novel protein, ECop (EGFR-Coamplified and overexpressed protein), in the regulation of NF-kappaB activity. Ectopic expression of ECop increases NF-kappaB transcriptional activity by promoting nuclear translocation and DNA binding of NF-kappaB, and ECop-induced NF-kappaB activation confers cellular resistance to apoptotic challenge. In ECop knockdown cells, NF-kappaB transcriptional activity is suppressed due to delayed IkappaBalpha degradation, which results in a delayed nuclear translocation as well as decreased DNA binding of NF-kappaB. Suppression of NF-kappaB activation by ECop knockdown increases cellular susceptibility to apoptosis. These results suggest that ECop is a key regulator of NF-kappaB signaling, and that high-level, amplification-mediated ECop expression, such as that occurring in tumors with amplified EGFR, could contribute to resistance to apoptosis.