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

Version 2 2019-11-07, 18:24
Version 1 2019-11-07, 17:26
Posted on 2019-11-07 - 18:24 authored by Amar Bhagwat

Assembly of fully infectious influenza A viruses (IAV) 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 volumes/s, 330 nm isotropic spatial resolution), we quantify Rab11A and vRNA movement in live cells during IAV infection. Here we report that IAV infection decreases the speed and increases the arrest of Rab11A. Unexpectedly, infection with respiratory syncytial virus (RSV) alters Rab11A motion in a manner opposite to IAV, 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 IAV vRNA in infected cells and provide direct evidence that vRNA-associated Rab11A have altered transport. The mechanism of altered Rab11A movement is likely related to a decrease in dynein motors bound to Rab11A vesicles during IAV infection.

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