Differential proteome analysis of host cells infected with porcine circovirus type 2.
Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome, which is an emerging swine immunosuppressive disease. To uncover cellular protein responses in PCV2-infected PK-15 cells, the comprehensive proteome profiles were analyzed utilizing two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF/TOF identification. Multiple comparisons of 2-DE revealed that the majority of changes in protein expression occurred at 48-96 h after PCV2 infection. A total of 34 host-encoded proteins, including 15 up-regulated and 19 down-regulated proteins, were identified by MALDI-TOF/TOF analysis. According to cellular function, the differential expression proteins could be sorted into several groups: cytoskeleton proteins, stress response, macromolecular biosynthesis, energy metabolism, ubiquitin-proteasome pathway, signal transduction, gene regulation. Western blot analysis demonstrated the changes of alpha tubulin, beta actin, and cytokeratin 8 during infection. Colocalization and coimmunoprecipitation analyses confirmed that the cellular alpha tubulin interacts with the Cap protein of PCV2 in the infected PK-15 cells. These identified cellular constituents have important implications for understanding the host interactions with PCV2 and brings us a step closer to defining the cellular requirements for the underlying mechanism of PCV2 replication and pathogenesis.
Zhang
Xin
X
Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, PR China.
Zhou
Jiyong
J
Wu
Yongping
Y
Zheng
Xiaojuan
X
Ma
Guangpeng
G
Wang
Zhongtian
Z
Jin
Yulan
Y
He
Jialing
J
Yan
Yan
Y
D000818:Animals; D018173:Circoviridae Infections / Q000378:metabolism*; D017925:Circovirus / Q000378:metabolism*; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction / Q000502:physiology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013552:Swine; D013553:Swine Diseases / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism
D020543:Proteome; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex