title: Label-free quantitative phosphoproteomic analysis reveals differentially regulated proteins and pathway in PRRSV-infected pulmonary alveolar macrophages.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen of swine worldwide and causes significant economic losses. Through regulating the host proteins phosphorylation, PRRSV was found to manipulate the activities of several signaling molecules to regulate innate immune responses. However, the role of protein phosphorylation during PRRSV infection and the signal pathways responsible for it are relatively unknown. Here liquid chromatography-tandem mass spectrometry for label-free quantitative phosphoproteomics was applied to systematically investigate the global phosphorylation events in PRRSV-infected pulmonary alveolar macrophages. In total, we identified 2125 unique phosphosites, of which the phosphorylation level of 292 phosphosites on 242 proteins and 373 phosphosites on 249 proteins was significantly altered at 12 and 36 h pi, respectively. The phosphoproteomics data were analyzed using ingenuity pathways analysis to identify defined canonical pathways and functional networks. Pathway analysis revealed that PRRSV-induced inflammatory cytokines production was probably due to the activation of mitogen-activated protein kinase and NF-κB signal pathway, which were regulated by several protein kinases during virus infection. Interacting network analysis indicated that altered phosphoproteins were involved in cellular assembly and organization, protein synthesis, molecular transport, and signal transduction in PRRSV infected cells. These pathways and functional networks analysis could provide direct insights into the biological significance of phosphorylation events modulated by PRRSV and may help us elucidate the pathogenic mechanisms of PRRSV infection.
authors.affiliation: Division of Animal Infectious Diseases, State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University , No. 1 Shi-zi-shan Street, Wuhan, Hubei 430070, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Luo
authors.forename: Liurong
authors.initials: L
authors.lastname: Fang
authors.forename: Hui
authors.initials: H
authors.lastname: Jin
authors.forename: Dang
authors.initials: D
authors.lastname: Wang
authors.forename: Kang
authors.initials: K
authors.lastname: An
authors.forename: Ningzhi
authors.initials: N
authors.lastname: Xu
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
authors.forename: Shaobo
authors.initials: S
authors.lastname: Xiao
mesh_terms: D000818:Animals; D016207:Cytokines / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D054884:Host-Pathogen Interactions; D016676:Macrophages, Alveolar / Q000737:chemistry* / Q000378:metabolism / Q000821:virology; D020928:Mitogen-Activated Protein Kinases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D058977:Molecular Sequence Annotation; D016328:NF-kappa B / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D019316:Porcine respiratory and reproductive syndrome virus / Q000502:physiology; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D013552:Swine
chemical_list: D016207:Cytokines; D016328:NF-kappa B; D010750:Phosphoproteins; D020928:Mitogen-Activated Protein Kinases