Classical swine fever virus down-regulates endothelial connexin 43 gap junctions.
Classical swine fever is a contagious disease of pigs characterized by fatal hemorrhagic fever. Classical swine fever virus (CSFV) induces the expression of pro-inflammatory and pro-coagulant factors of vascular endothelial cells and establishes a long-term infection. This study aimed to understand the effect of CSFV on endothelial connexin 43 (Cx43) expression and gap junctional intercellular coupling (GJIC). Porcine aortic endothelial cells were infected with CSFV at different multiplicity of infection for 48 h. Semi-quantitative RT-PCR, immunoconfocal microscopy, and Western blotting showed that the transcription and translation of Cx43 were reduced, and this was associated with an attenuation of GJIC. This decrease occurred in a time-dependent manner. An ERK inhibitor (PD98059), a JNK inhibitor (SP600125), and proteasome/lysosome inhibitors all significantly reversed the reduction in Cx43 protein levels without any influence on the titer of progeny virus. In addition, CSFV activated ERK and JNK in a time-dependent manner and down-regulated Cx43 promoter activity, mainly through decreased AP2 binding. This effect was primarily caused by the replication of CSFV rather than a consequence of cytokines being induced by CSFV infection of endothelial cells.
Hsiao
Hsiang-Jung
HJ
Department of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan.
Liu
Pei-An
PA
Yeh
Hung-I
HI
Wang
Chi-Young
CY
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D006692:Classical Swine Fever Virus / Q000502:physiology*; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D017629:Gap Junctions / Q000378:metabolism*; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors; D011401:Promoter Regions, Genetic; D013552:Swine; D014779:Virus Replication / Q000502:physiology
D018031:Connexin 43; D048049:Extracellular Signal-Regulated MAP Kinases; D048670:MAP Kinase Kinase 4