|The Herpesviridae family of DNA viruses is divided into the alpha, beta and gamma subfamilies that are represented by herpes simplex virus type 1 (HSV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) respectively. Successful herpesvirus replication involves alterations of many host cell pathways including cell cycle progression, DNA damage responses and apoptosis, which result from the actions of multiple viral proteins. During the course of infection, herpesviruses expresses between 80 and 200 viral proteins, many of which are poorly or completely uncharacterized. We have generated an expression library for most of the HSV, CMV and EBV proteins, and used it to determine the subcellular localization of these proteins and their effects on PML nuclear bodies. This led to the identification of several proteins from each virus that alter or disrupt host PML nuclear bodies, including the CMV UL35 protein, which we further characterized. Since lytic infection by herpesviruses is known to block cells at the G1/S phase of the cell cycle, we recently used the viral library to identify proteins that promote G1/S accumulation. Several proteins with this property were identified from each virus and their mechanisms of action can now be pursued. The viral library will continue to serve as a valuable tool for identifying viral proteins that manipulate specific host processes. The mechanisms of action of these viral proteins can then be examined using proteomic methods for identifying host protein interactions and effects on the host proteome using methods such as 2D Difference Gel Electrophoresis (DiGE). We recently used 2D-DiGE to identify changes in the nuclear proteome caused by EBNA1.|
Salamun, S.G., Sitz, J., De La Cruz-Herrera, C.F., Marcon, E., Greenblatt, J., Fradet-Turcotte, A. and Frappier, L. 2019 The Epstein-Barr Virus BMRF1 Protein Activates Transcription and Inhibits the DNA Damage Response by Binding NuRD. J. Virol. In press.
Paladino, P., Marcon, E., Greenblatt, J. and Frappier, L. 2014 Identification of Herpesvirus Proteins that Contribute to G1/S Arrest. J. Virol. 88, 4480-4492.
Salsman, J., Jagannathan, M., Paladino, P., Chan, P.-K., Dellaire, G., Raught, B. and Frappier, L. 2012 Proteomic Profiling of the Human Cytomegalovirus UL35 Gene Products Reveals a Role for UL35 in the DNA Repair Response. J. Virol. 86, 806-820. PMID: 22072767 (cover for vol 86 issue 4)
Cao, J. Y., Mansouri, S. and Frappier, L. 2012 Changes in the nasopharyngeal carcinoma nuclear proteome induced by the EBNA1 protein of Epstein-Barr virus reveal potential roles for EBNA1 in metastasis and oxidative stress responses. J. Virol. 86, 382-394. PMID: 22013061
Salsman, J., Zimmerman, N., Chen, T., Domagala, M. and Frappier, L. 2008 Genome-wide Screening of Herpes Simplex virus, Cytomegalovirus and Epstein-Barr virus Proteins for Subcellular Localization and Alteration of PML Nuclear Bodies. PLoS Pathogens 4(7): e1000100. doi:10.1371/journal.ppat.1000100
Maxwell, K.L. and Frappier, L 2007 Viral Proteomics. Microbiology and Molecular Biology Reviews, 71(2), 398-411.
Gao, M., Brufatto, N., Chen, T., Murley, L.L., Thalakada, R., Domagala, M., Beattie, B., Mamelak, D., Athanasopoulos, V., Johnson, D., McFadden, G., Burks, C. and Frappier, L. 2005 Expression Profiling of Herpesvirus and Vaccinia Virus Proteins using a High-Throughput Baculovirus Screening System. J. Proteome Research 4, 2225-2235. PMID: 16335970
Holowaty, M.N., Zeghouf, M., Wu, H., Tellam, J., Athanasopoulos, V., Greenblatt, J. and Frappier, L. 2003. Protein profiling with Epstein-Barr nuclear antigen 1 reveals an interaction with the herpesvirus associated ubiquitin specific protease, HAUSP/USP7. J.Biol. Chem. 278, 29987-29994. PMID: 12783858