Pathogenic negative strand RNA virus such as Ebola virus, measles virus, RSV and mumps virus derive their membrane envelope from the host cell. Infected cells produce new virus particles, which are thought to assemble and bud off the plasma membrane. Viral matrix proteins have been shown to be instrumental in this process and it is thought that multiple macromolecular interactions between the matrix protein and membranes, the nucleoprotein, which packages the RNA genome into ribo-nucleoprotein particles (RNP) and the cytoplasmic tails of viral glycoproteins act together to drive assembly and budding. There is also amounting evidence that enveloped viruses such as filo viruses (Ebola) and retroviruses (HIV) utilize the cellular protein machinery implicated in the formation of, multivesicular bodies, a process, which resembles virus budding morphologically. The aim of the proposal described here is to better understand the structural requirements of virus assembly, which may ultimately lead to the identification of new targets for anti-viral agents. We propose to solve the three dimensional structures of two Ebola virus matrix protein complexes:
(i)the matrix protein in complex with the cellular budding factor Tsg 101 and
(ii) the matrix protein in complex with a fragment of the nucleoprotein. The elucidation of the molecular interactions will give insight into the assembly and budding mechanism of other pathogenic negative strand RNA viruses. The applicant will enlarge and strengthen her background in protein chemistry. This comprises general protein chemistry techniques, including expression studies protein complex formation and characterization; important skills, which are instrumental for a scientist in the post genomic, are. Furthermore, the applicant will have the opportunity to learn protein crystallography and acquire a strong and solid expertise in the field of structural biology. In return, the applicants experience in molecular biology will benefit all other members of the host laboratory.