Proteomic and Phosphoproteomic Analysis of Chicken Embryo Fibroblasts Infected with Cell Culture-Attenuated and Vaccine Strains of Marek’s Disease Virus

Abstract

Vaccination is an effective strategy to reduce the loss of chickens in the poultry industry caused by Marek's Disease (MD), an avian lymphoproliferative disease. The vaccines currently used are from attenuated serotype 1 Marek's disease virus (MDV) or naturally nononcogenic MDV strains. To prepare for future immunity breaks, functional genomic and proteomic studies have been used to better understand the underlying mechanisms of MDV pathogenicity and the effects induced by the vaccine viruses. In this study, a combined approach of quantitative GeLC-MSE and qualitative ERLIC/IMAC/LC-MS/MS analysis were used to identify abundance changes of proteins and the variations of phosphorylation status resulting from the perturbations due to infection with an attenuated oncogenic virus strain (Md11/75C) and several nononcogenic virus strains (CVI988, FC126 and 301B) in vitro. Using this combined approach, several signal transduction pathways mapped by the identified proteins were found to be altered at both the level of protein abundance and phosphorylation. On the basis of this study, a kinase-dependent pathway to regulate phosphorylation of 4E-BP1 to modulate assembly of the protein translation initiation complex was revealed. The differences of 4E-BP1 phosphorylation patterns as well as the measured abundance changes among several other proteins that regulate host transcriptional and translational activities across the virus strains used in this study provide new insight for future functional and biochemical characterization of specific proteins involved in MDV pathogenesis.

Proteomics of Breast Muscle Tissue Associated with the Phenotypic Expression of Feed Efficiency within a Pedigree Male Broiler Line: I. Highlight on Mitochondria

“…Proteomic approaches are promising as they can circumvent some of the issues associated with transcriptomics. Various proteomic studies in poultry have recently emerged including reports on muscle [ 16 , 17 ], eggs [ 18 – 20 ], and liver [ 21 ] as well as host-virus interactions [ 22 – 26 ]. Muscle proteomics in livestock has been extensively studied for physiological relevance associated with specific diseases, metabolic conditions, and production traits (See review by Picard et al [ 27 ]).…”

“…Advances in all areas of the MS-based proteomics workflow, including sample preparation, liquid chromatography (LC)-MS and computational analysis, have made it a preferred analytic tool to characterize protein dynamics in multiple functional dimensions; and proteomics based on LC-MS has essentially replaced previous tools such as 2DGE. The shotgun proteomics method has been used in poultry [ 22 ].…”

Growth Factor Priming Differentially Modulates Components of the Extracellular Matrix Proteome in Chondrocytes and Synovium-Derived Stem Cells

“…The combination of label-free MS E data acquisition and post-processing with Elucidator data mining software for accurate mass and retention time matching is validated independently by other groups [35], [39]–[41]. From one C-P sample, all three chromatograms were outliers (derived from this one sample), were excluded from the dataset based as typical on visual inspection [42] and principal component analysis (data not shown).…”

New Approaches to Comparative and Animal Stress Biology Research in the Post-genomic Era: A Contextual Overview

“…A few examples of studies exploring phospho-proteomics in non-model organisms include phospho-proteome of chicken ( Gallus gallus ) embryo fibroblasts and of the mitochondria of hibernating thirteen-lined ground squirrels ( I . tridecemlineatus ) [64,65] . Recently, studies are beginning to explore the methyl-proteome by the enrichment of methylated protein through pan-methyl-arginine antibodies or methyl-lysine binding domains [66] .…”