2012 Annual Report
1a.Objectives (from AD-416):
The objective of this collaborative research project is to identify the host proteins that play critical roles in the process of virus replication. Identified host proteins will be characterized in terms of cell location, interaction’s kinetics, and participation of the virus cycle. Additionally, the mapping of the specific areas of the host proteins responsible of the interaction with the virus proteins will be identified.
1b.Approach (from AD-416):
The identified host proteins will be identified by the two hybrid yeast system. Those identified will be further characterized, particularly the molecular interaction between the host and virus proteins. Residues in the host proteins mediating the interaction with the virus proteins will be identified and characterized. Based on that information, the role of those residues in the process of virus replication will be assessed. Monoclonal antibodies specific for the different virus proteins are critical tools for this type of study since they allow the identification of the virus proteins in different methodological procedures to study the characteristics of the interaction between the host and virus proteins.
The goal of this collaboration is to utilize a set of monoclonal antibodies recognizing virus proteins specific to Foot-and-Mouth Disease Virus (FMDV). These proteins will aid in the development of novel therapeutic methodologies to control or abort FMDV replication.
During FY 2012, the host proteins that specifically interact with virus proteins were identified in bovine cells infected with FMDV. Specifically, several bovine proteins, including Beclin1, which is an important cell factor involved in intracellular traffic and degradation of cellular proteins, bind non structural protein 2C. The nature of the reactivity was characterized and its importance in virus replication analyzed. Monoclonal antibodies against FMDV 2C were used to study the interaction between the host and virus proteins and individualize the viral protein into the infected cells at different stages of the interaction with 2C. A manuscript entitled, “Foot and Mouth Disease Virus non structural protein 2C interacts with Beclin1 modulating virus replication” by D. P. Gladue, V. O’Donnell, R. Baker-Branstetter, J.M. Pacheco-Tobin, L.G. Holinka, I. Fernandez Sainz, Z. Lu, J. Zhu, M. Puckette, E. Brocchi, L. Rodriguez, and M. V. Borca was submitted to Journal of Virology.
The deliverables of this collaborative agreement compliment ARS in-house research project 1940-32000-057-00D, Intervention strategies to support the global control and eradication of foot-and-mouth disease virus, objective 1 Elucidate the host-pathogen interaction of FMDV.