Abstract
ABSTRACT Classical swine fever virus (CSFV) is one of the most important viral pathogens leading worldwide threats to pig industry. MicroRNAs (miRNAs) play important roles in regulating virus replication, but whether miRNAs affect CSFV infection is still poorly understood. In previous study, we identified four miRNAs that were down-regulated by CSFV in swine umbilical vein endothelial cells (SUVEC). In this study, miR-140, one of the most potently down-regulated genes was investigated. We found that the miRNA expression was significantly inhibited by CSFV infection. Subsequent studies revealed that miR-140 mimics significantly inhibited CSFV replication, while the inhibition of endogenous miR-140 enhanced CSFV replication. By using bioinformatics prediction, luciferase reporter system, real-time fluorescence quantitative PCR (RT-qPCR) and Western blot assays, we further demonstrated that miR-140 bind to the 3ʹ UTR of Rab25 mRNA to regulate its expression. We also analyzed the expression pattern of Rab25 in SUVECs after CSFV infection. The results showed that CSFV infection induced Rab25 expression. Finally, Rab25 was found to promote CSFV replication. In conclusion, this study demonstrated that CSFV inhibits miR-140 expression and miR-140 inhibits replication by binding to host factor Rab25.
Highlights
Classical swine fever virus (CSFV) is an important member of pestivirus genus within the Flaviviridae family [1]
The results showed that CSFV replication reached the platform stage at 48 h post-infection (Figure 1(b))
swine umbilical vein endothelial cells (SUVEC) were infected with CSFV at a 0.1 multiplicity of infection (MOI) and the miR-140 expression level was measured at different time postinfection
Summary
Classical swine fever virus (CSFV) is an important member of pestivirus genus within the Flaviviridae family [1]. Classical swine fever (CSF) is one of the diseases notifiable to the World Organization for Animal Health and represents one of the leading worldwide threats to pig industry. Persistent CSFV infection is the leading cause of chronic CSF. Elucidate the pathogenic mechanism of CSFV may largely help to develop effective prevention and control strategies. Rab GTPases comprise the largest subfamily of small GTPases and play a master role in regulating intercellular vesicle and protein transport [2]. The Ras small G protein superfamily have been identified as a key regulator of the intracellular transport system [4]. Rab is involved in transcytosis, endocytic sorting, and transport [5]. It has been reported that the aberrant expression of Rab was linked to cancer development and Rab is recognized as an oncogene [6]
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