Ebola is a negative stranded RNA virus. Its pathogenesis is characterized by internal and external bleeding in humans and other primates as a result of coagulation abnormalities induced by the virus at the onset of the infection. Since its first discovery in 1976, no specific treatment or vaccines have been found. It is classified as biosafety level IV agent and therefore has the potential to be weaponised. The mechanistic details of the virus assembly process are lacking. Evidence suggests that the two main matrix proteins VP 40 and VP24 play crucial role in the budding of the virus. Generation of new virus involve a cascade of cellular events that recruits the viral genome, the matrix proteins and subsequent acquisition of the viral envelope from the host cell. The virus like particle (VLP) is extruded from the host cell through a mechanism that is not fully understood. This can serve as a primary target for Ebola virus replication. Available data suggest that VP 40 and 24 bind lipid membranes with very high affinity and alter membrane curvature, a process which is crucial for the egress of the virus. In this proposed research we seek to elucidate the mechanistic details of lipid interaction by Ebola matrix proteins and their mechanism of generating new viral particles. Specifically, we seek to determine 1) the membrane targeting and curvature alteration mechanisms of VP40 and VP24. 2) the interplay of F-actin targeting of V24 and the ability to alter membrane curvature. 3) the lipid dependent interaction properties of VP40 and human Nedd4 in the generation of Ebola VLPs. Methodologies to be employed in the studies include a) an array of biophysical methods to analyze the interaction of matrix proteins with various membrane models. These include: monolayer, surface plasmon resonance, sedimentation and stop-flow analysis. b) Cellular targeting and trafficking of the matrix proteins using fluorescent tags. c) Structural investigation of membrane curvature process with X-ray and fluorescent methods. c) F-actin binding properties of VP24. Given the dearth of mechanistic details about Ebola replication, this studies is expected to elucidate the mechanism by which the Ebola virus interacts with the host plasma membrane and how new viruses are generated. This will provide clues to the development of therapeutic protocols for Ebola hemorrhagic virus.