Foot-and-mouth disease virus modulates cellular vimentin for virus survival

Authors: GLADUE, DOUGLAS - University Of Connecticut; O'DONNELL, VIVIAN - University Of Connecticut; BAKER-BRANSTETTER, RYAN - Oak Ridge Institute For Science And Education (ORISE); Holinka-Patterson, Lauren; Pacheco Tobin, Juan; Fernandez Sainz, Ignacio; LU, ZHIQIANG - Us Deparment Of Homeland Security; BROCCHI, E - Lombardy And Emilia Romagna Experimental Zootechnic Institute; PICCONE, MARIA - University Of Connecticut; FLETCHER, PAIGE - Oak Ridge Institute For Science And Education (ORISE); AMBROGGIO, XAVIER - National Institutes Of Health (NIH); Rodriguez, Luis; Borca, Manuel

Submitted to: Journal of Virology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/29/2013
Publication Date: 4/10/2013
Publication URL: http://handle.nal.usda.gov/10113/57143
Citation: Gladue, D.P., O'Donnell, V., Baker-Branstetter, R., Holinka-Patterson, L.G., Pacheco Tobin, J., Fernandez Sainz, I.J., Lu, Z., Brocchi, E., Piccone, M.E., Fletcher, P., Ambroggio, X., Rodriguez, L.L., Borca, M.V. 2013. Foot-and-mouth disease virus modulates cellular vimentin for virus survival. Journal of Virology. 87(12):6794-6803.

Interpretive Summary: Foot-and-mouth disease virus (FMDV) is the causative agent of foot-and-mouth disease. During infection with FMDV, the cell presents several areas where virus replication takes place. In these areas one of the virus proteins, 2C, appears to have multiple roles during virus replication. To better understand the role of 2C in the process of virus replication, we have been using a specific methodology, yeast two-hybrid, to identify cell host proteins that interact with virus 2C. Here, we report that a cellular protein, vimentin, specifically interacts with 2C. The 2C-vimentin interaction was confirmed to occur in cells infected with FMDV by using two different independent additional methodologies (co-immunoprecipitation and fluorescent microscopy). Manipulation of intracellular levels of vimentin showed that reduced levels of vimentin resulted in a significant decrease in viral yield, indicating that vimentin plays an important role in the replication cycle of FMDV. The use of Acrylamide, which causes disruption of intracellular vimentin filaments, also inhibited viral yield in FMDV infected cells. We also identified the areas of 2C being critical for vimentin binding. It also showed that the integrity of those areas in 2C are necessary for virus growth, suggesting that the interaction between FMDV 2C and cellular vimentin is essential for virus replication.

Technical Abstract: Foot-and-mouth disease virus (FMDV), the causative agent of foot-and-mouth disease, is an Apthovirus within the Picornaviridae family. During infection with FMDV, several host cell membrane rearrangements occur to form sites of viral replication. The largest viral protein in the replication complex, 2C, is thought to have multiple roles during virus replication. To better understand the role of 2C in the process of virus replication, we have been using a yeast two-hybrid approach to identify host proteins that interact with 2C. We recently reported that cellular Beclin1 is a natural ligand of 2C and that it is involved in the autophagy pathway which was shown to be important for FMDV replication. Here, we report that cellular vimentin is also a specific host binding partner for 2C. The 2C-vimentin interaction was further confirmed by co-immunoprecipitation and deconvolution microscopy to occur in FMDV-infected cells. It was shown that upon infection a vimentin structure forms around 2C, and that this structure is later resolved or disappears. Interestingly, over-expression of vimentin had no effect on virus replication; however, over-expression of a truncated dominant-negative form of vimentin resulted in a significant decrease in viral yield. Acrylamide, which causes disruption of vimentin filaments, also inhibited viral yield. Alanine scanning mutagenesis was used to map the specific amino acid residues in 2C critical for vimentin binding. Using reverse genetics, we identified 2C residues that are necessary for virus growth, suggesting that the interaction between FMDV 2C and cellular vimentin is essential for virus replication.