Foot-and-mouth disease virus replication sites form next to the nucleus and close to the Golgi apparatus, but exclude marker proteins associated with host membrane compartments.
Picornavirus infection of cells generally results in the production of membranous vesicles containing the viral proteins necessary for viral RNA synthesis. To determine whether foot-and-mouth disease virus (FMDV) infection induced similar structures, and which cellular components were involved, the subcellular distribution of FMDV proteins was compared with protein markers of cellular membrane compartments. Using immunofluorescence analysis and digital deconvolution, it was shown that FMDV structural and non-structural proteins co-localize to punctate structures in juxtanuclear virus assembly sites close to the Golgi complex. Significantly, viral protein 2C did not co-localize with marker proteins of the cis- or medial-Golgi compartments or trans-Golgi network. Furthermore, incubation of infected cells with brefeldin A caused a redistribution of Golgi proteins to the endoplasmic reticulum, but did not affect the distribution of 2C and, by inference, the integrity of the virus assembly site. Taken with the observation that 2C was membrane-associated, but failed to fractionate with Golgi markers on density gradients, it was possible to conclude that Golgi membranes were not a source of structures containing 2C. Further immunofluorescence analysis showed that 2C was also separate from marker proteins of the endoplasmic reticulum, endoplasmic reticulum intermediate compartment, endosomes and lysosomes. The results suggest that the membranes generated at FMDV assembly sites are able to exclude organelle-specific marker proteins, or that FMDV uses an alternative source of membranes as a platform for assembly and replication.
Knox
Caroline
C
University of St Andrews, School of Biology, Centre for Biomolecular Sciences, Biomolecular Sciences Building, North Haugh, St Andrews KY16 9ST, UK.
Moffat
Katy
K
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Surrey GU24 0NF, UK.
Ali
Shireen
S
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Surrey GU24 0NF, UK.
Ryan
Martin
M
University of St Andrews, School of Biology, Centre for Biomolecular Sciences, Biomolecular Sciences Building, North Haugh, St Andrews KY16 9ST, UK.
Wileman
Thomas
T
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Surrey GU24 0NF, UK.
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D029544:Foot-and-Mouth Disease Virus / Q000502:physiology*; D006056:Golgi Apparatus / Q000502:physiology; D007425:Intracellular Membranes / Q000378:metabolism* / Q000821:virology; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
D014764:Viral Proteins