title: Understanding Epstein-Barr Virus Life Cycle with Proteomics: A Temporal Analysis of Ubiquitination During Virus Reactivation.
abstract: Epstein-Barr virus (EBV) is a human γ-herpesvirus associated with cancer, including Burkitt lymphoma, nasopharyngeal, and gastric carcinoma. EBV reactivation in latently infected B cells is essential for persistent infection whereby B cell receptor (BCR) activation is a physiologically relevant stimulus. Yet, a global view of BCR activation-regulated protein ubiquitination is lacking when EBV is actively replicating. We report here, for the first time, the long-term effects of IgG cross-linking-regulated protein ubiquitination and offer a basis for dissecting the cellular environment during the course of EBV lytic replication. Using the Akata-BX1 (EBV+) and Akata-4E3 (EBV-) Burkitt lymphoma cells, we monitored the dynamic changes in protein ubiquitination using quantitative proteomics. We observed temporal alterations in the level of ubiquitination at ∼150 sites in both EBV+and EBV-B cells post-IgG cross-linking, compared with controls with no cross-linking. The majority of protein ubiquitination was downregulated. The upregulated ubiquitination events were associated with proteins involved in RNA processing. Among the downregulated ubiquitination events were proteins involved in apoptosis, ubiquitination, and DNA repair. These comparative and quantitative proteomic observations represent the first analysis on the effects of IgG cross-linking at later time points when the majority of EBV genes are expressed and the viral genome is actively being replicated. In all, these data enhance our understanding of mechanistic linkages connecting protein ubiquitination, RNA processing, apoptosis, and the EBV life cycle.
authors.affiliation: 1 Philips Institute for Oral Health Research, VCU School of Dentistry, Virginia Commonwealth University , Richmond, Virginia.
authors.forename: Dong-Wen
authors.initials: DW
authors.lastname: Lv
authors.affiliation: 2 McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine , Baltimore, Maryland.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhong
authors.affiliation: 1 Philips Institute for Oral Health Research, VCU School of Dentistry, Virginia Commonwealth University , Richmond, Virginia.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhang
authors.affiliation: 2 McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine , Baltimore, Maryland.|3 Institute of Bioinformatics , International Technology Park, Bangalore, India .|4 Diana Helis Henry Medical Research Foundation , New Orleans, Louisiana.
authors.forename: Akhilesh
authors.initials: A
authors.lastname: Pandey
authors.affiliation: 1 Philips Institute for Oral Health Research, VCU School of Dentistry, Virginia Commonwealth University , Richmond, Virginia.|5 Department of Microbiology and Immunology, School of Medicine, Virginia Commonwealth University , Richmond, Virginia.|6 Massey Cancer Center, Virginia Commonwealth University , Richmond, Virginia.
authors.forename: Renfeng
authors.initials: R
authors.lastname: Li
mesh_terms: D001402:B-Lymphocytes / Q000378:metabolism; D045744:Cell Line, Tumor; D004854:Herpesvirus 4, Human / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D040901:Proteomics / Q000379:methods*; D054875:Ubiquitination
keywords: Association Study; big data; proteomics