Ebola virus (EBOV) VP24, together with nucleoprotein and VP35, is an essential component of viral RNA-protein complexes called "nucleocapsids." In this study, using a series of deletion mutants of VP24, we identified regions within VP24 that are important for the formation of nucleocapsid-like structures and determined that both termini of VP24 are essential for nucleocapsid formation. This finding advances our knowledge of both EBOV morphogenesis and the nature of VP24 molecules in nucleocapsid formation, which will be useful for the development of antiviral compounds.