Visualization and formation West Nile Virus replication complexes
During WNV replication, intracellular membrane structures are dramatically altered. Towards the end of the latent period, translation is observed in virally induced membrane structures called convoluted membranes (CM) and paracrystalline arrays (PC) and RNA synthesis is observed in vesicle packets (VP).These membrane structures are crucial for the efficient replication of flaviviruses and are linked to the exponential increase in virus production. However, previous visualization of these structures has been static at fixed time points. We aim to visualize the formation and proliferation of these virally induced membrane structures over real time using time-lapse epi-fluorescence. Recently, proteins localizing to the CM/PC and VP have been identified and we aim to tag these proteins, in frame, with a reporter protein to visualize WNV replication over real-time. We are also interested in determining host/viral protein interactions for membrane biogenesis. A number of host proteins are known to form cellular membrane structure. During infection, these cellular proteins can be recruited by viral proteins to aid in CM/PC and VP formation. Identifying and understanding these interactions can allow us to determine WNV replication and which components/organelles are used and abused during viral infection.
Mr Turgut Aktepe (PhD Student)
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