Expression of survivin using SFV and analysis of phosphorylation site mutagenesis.

Apoptosis is a natural process, essential to the development and homeostatis of the organism, in which a cell undergoes self-mediated death. The human inhibitor of apoptosis (IAP) survivin was previously characterized by its role in the pathogenesis of various cancers. Although most adult tissues do not contain measurable amounts of survivin transcript or protein, it is up-regulated in many cancers. AstraZeneca is currently focused on developing small molecule therapies to target survivin in vivo. In this project a Semliki Forest Virus (SFV) system was used to generate survivin in its native form. Western analysis of transduced HeLa and BHK-21 cells showed no visible survivin production. Additionally, the effects of phosphorylation on survivin\u27s function in cell cycle checkpoint were analyzed using point mutation constructs. Western analysisof tranfected HeLa cells with these constructs showed no visible survivin production, and immunocytochemical analysis showed no effect on cell cycle checkpoint