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Quantitative Live Cell Imaging of Rab11A and Viral RNA Reveals Differential Manipulation of Transport Pathways

Assembly of fully infectious influenza viruses is a complex process involving transport from the nucleus to the plasma membrane. Rab11A containing recycling endosomes have been identified as a platform for intracellular transport of viral RNA (vRNA). Using high spatiotemporal resolution light-sheet microscopy (~1.4 cell volumes/s, 330 nm isotropic spatial resolution), we quantify Rab11A and vRNA movement in live-cells during influenza virus infection. We demonstrate that influenza virus infection decreases the speed and increases the arrest of Rab11A. 

Unexpectedly, infection with RSV alters Rab11A motion in a manner opposite to influenza virus, suggesting that Rab11A is a common host component that is differentially manipulated by respiratory RNA viruses. Using two-color imaging we demonstrate co-transport between Rab11A and vRNA in infected cells and provide direct evidence that vRNA-associated Rab11A have altered transport. The mechanism of altered Rab11A movement is likely due to a decrease in dynein motors bound to Rab11A vesicles during influenza virus infection.

Dr Seema Lakdawala, Lakdawala Lab, University of Pittsburgh

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