Abstract
Within 2–6 hours after infection by vesicular stomatitis virus (VSV), newly assembled VSV particles are released from the surface of infected cells. In that time, viral ribonucleoprotein (RNP) particles (nucleocapsids) travel from their initial sites of synthesis near the nucleus to the edge of the cell, a distance of 5–10 μm. The hydrodynamic radius of RNP particles (86 nm) precludes simple diffusion through the mesh of cytoskeletal fibers. To reveal the relative importance of different transport mechanisms, movement of GFP-labeled RNP particles in live A549 cells was recorded within 3 to 4 h postinfection at 100 frames/s by fluorescence video microscopy. Analysis of more than 200 RNP particle tracks by Bayesian pattern recognition software found that 3% of particles showed rapid, directional motion at about 1 μm/s, as previously reported. 97% of the RNP particles jiggled within a small, approximately circular area with Gaussian width σ = 0.06 μm. Motion within such “traps” was not directional. Particles stayed in traps for approximately 1 s, then hopped to adjacent traps whose centers were displaced by approximately 0.17 μm. Because hopping occurred much more frequently than directional motion, overall transport of RNP particles was dominated by hopping over the time interval of these experiments.
Highlights
Within 2–6 hours after infection by vesicular stomatitis virus (VSV), newly assembled VSV particles are released from the surface of infected cells
For particles too large to diffuse in the cytoplasm[2], movements consist of a mixture of seemingly random motions and directional motion driven by molecular motors on microtubules and actin filaments[3]
RNP particles were imaged in cells infected with recombinant VSV containing a gene for enhanced green fluorescent protein fused in frame with the P gene (VSV-P-eGFP)[8]
Summary
Within 2–6 hours after infection by vesicular stomatitis virus (VSV), newly assembled VSV particles are released from the surface of infected cells. For particles too large to diffuse in the cytoplasm (hydrodynamic radius> approximately 50 nm)[2], movements consist of a mixture of seemingly random motions and directional motion driven by molecular motors on microtubules and actin filaments[3]. The purpose of the experiments presented here was to determine the transport modes of the ribonucleoprotein (RNP) core (nucleocapsid) of vesicular stomatitis virus (VSV) across the cytoplasm using a variational Bayesian approach to analyze single particle tracks in living cells[4]. Vesiculovirus Indiana, commonly referred to as VSV (Indiana serotype), is one of the prototypes for the large group of viruses with nonsegmented negative strand RNA genomes (order Mononegavirales) These viruses include many human pathogens, such as measles, Ebola, and rabies viruses, as well as many other viruses that infect both animals and plants. Our limited data suggests that driven motion is sometimes directed away from the nucleus and sometimes directed toward the nucleus, undercutting this argument
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