Molecular cloning and characterization of amida, a novel protein which interacts with a neuron-specific immediate early gene product arc, contains novel nuclear localization signals, and causes cell death in cultured cells

Amida was isolated by the yeast two-hybrid system as a novel protein which associated with Are, a non-transcriptional immediate early gene specific to the brain. Amida was confirmed to be associated with Are in vitro and in v ivo. Amida shows no homology to known proteins. Amida is ubiquitously expre ssed, although it is abundant in the brain. A transfection study revealed t hat Amida was localized in the nucleus and after 72 h the transfected cells underwent apoptosis, Furthermore, we found two nuclear localization signal s and a domain needed for interacting with Are was encompassed by two nucle ar localization signals. Co-transfection experiment with Amida and Are sugg ested that Amida transported Are into the nucleus and negatively regulated Amida-induced cell death. These results indicate that Are together with Ami da may modulate cell death in the brain.