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NJIT Mathematical Biology Seminar

Tuesday, January 22, 2008, 4:00pm
Cullimore Hall 611
New Jersey Institute of Technology

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Arf1 Dynamics in Coxsackievirus Infected cells

Ani Chintalapani & Matthew Hanna

Undergraduate Biology and Math Training Program, NJIT

Coxsackie virus Type 3 (CVB3) like other entroviruses are single (+) stranded RNA viruses which use the cytoplasmic surface of intracellular membranes for RNA replication. Numerous studies have shown that Coxsackie virus replication is sensitive to the fungal toxin Brefeldin A (BFA), a well-characterized, highly specific inhibitor of Arf1 GTPase activation, but the capacity in which Arf1 assists the RNA replication is not understood. In uninfected cells active GTP-bound state Arf1 associates with nascent endoplasmic reticulum (ER) export domains, recruiting effectors whose activities, ranging from regulating membrane curvature and cytoskeletal machinery to signaling, result in the biogenesis of the ER-Golgi Intermediate Compartment (ERGIC) membranes which bud off and fuse with the Golgi apparatus. Given this, Arf1activity in infected cells may also be utilized to specialized unconventional organelles for RNA replication. We have previously shown that in CVB3 infected cells Arf1 accumulates on discrete domains of the endoplasmic reticulum membranes which colocalize with viral replication machinery components. Here using fluorescence recovery after photobleaching techniques, we investigate the changes in Arf1 membrane binding and dissociation rates in single living cells at different time points during infection. With these rates and our measurements of the abundances of Arf1 at these accumulated domains, we generate a model of Arf1 dynamics in CVB3 infected cells.