Vaccinia virus cores are transported on microtubules

Abstract

Infection with Vaccinia virus (VV) produces several distinct virions called intracellular mature virus (IMV), intracellular enveloped virus (IEV), cell-associated enveloped virus (CEV) and extracellular enveloped virus (EEV). In this report, we have investigated how incoming virus cores derived from IMV are transported within the cell. To do this, recombinant VVs (vA5L-EGFP-N and vA5L-EGFP-C) were generated in which the A5L virus core protein was fused with the enhanced green fluorescent protein (EGFP) at the N or C terminus. These viruses were viable, induced formation of actin tails and had a plaque size similar to wild-type. Immunoblotting showed the A5L-EGFP fusion protein was present in IMV particles and immunoelectron microscopy showed that the fusion protein was incorporated into VV cores. IMV made by vA5L-EGFP-N were used to follow the location and movement of cores after infection of PtK 2 cells. Confocal microscopy showed that virus cores were stained with anti-core antibody only after they had entered the cell and, once intracellular, were negative for the IMV surface protein D8L. These cores co-localized with microtubules and moved in a stop-start manner with an average speed of 51?8 (±3?9) mm min "1 , consistent with microtubular movement. Treatment of cells with nocodazole or colchicine inhibited core movement, but addition of cytochalasin D did not. These data show that VV cores derived from IMV use microtubules for intracellular transport after entry.

Identification of new members of Fertilisation Independent Seed Polycomb Group pathway involved in the control of seed development inArabidopsis thaliana

“…Potential candidates are microtubules that have been reported to be organised in an orientation compatible with their use as tracks for migration of NCDs (Brown and Lemmon, 2001). However, the speed of migration of NCDs is at least 50 times slower than the speed recorded for migration of organelles along microtubules (Pollock et al, 1998;Carter et al, 2003). Alternatively, actin filaments may be suitable candidates, although no actin cable was reported at the posterior pole of endosperm in Coronopus didymus, a relative of Arabidopsis (Nguyen et al, 2002).…”

Synergistic anti-tumor effects between oncolytic vaccinia virus and paclitaxel are mediated by the IFN response and HMGB1

“…19 We used a vaccinia strain with Green Fluorescent Protein (GFP) fused to the viral A5L protein, a DNA packaging protein that is one of the most abundant within the viral particle. 20 As such, it is possible to detect single viral particles by fluorescence microscopy or flow cytometry. This virus was used along with 7-amino-actinomycin staining to assess the relative levels of initial infection and the position in the cell cycle of HCT-116 cells alone or following paclitaxel treatment (as determined 2 h after addition of the virus, before the viral infection itself can affect the position of the cells in the cell cycle).…”

“…35 The WR strain containing an A5L-GFP fusion was provided by Geoff Smith (Imperial College, London, UK). 20 …”

Electron microscopic examination of the viromatrix of Rana grylio virus in a fish cell line

“…The morphogenesis of RGV showed that virus particles assembled in the viromatrix (Zhang et al, 1999(Zhang et al, , 2001(Zhang et al, , 2002. Studying the ultrastructure of viromatrix contributes to understanding the assembly pathway and pathogenesis (Granzow et al, 1997;Wolf et al, 2000;Griffiths et al, 2001;Qin et al, 2001;Chao et al, 2004), and the relationships between the viromatrix and cellular components, such as the cytoskeleton (Hollinshead et al, 2001;Carter et al, 2003), mitochondria (Rojo et al, 1998) and the endoplasmic reticulum (ER) (Sodeik et al, 1993;Risco et al, 2002), have provided impor-tant insights into virus assembly. The viromatrix of large DNA viruses had been investigated in detail by electron microscopy, for example, African swine fever virus (ASFV) and vaccinia virus (VV) (Andres et al, 1998;Rouiller et al, 1998;Griffiths et al, 2001).…”