Abstract
Cellular levels of phosphatidylinositol 4-phosphate (PI4P) have been shown to be upregulated during RNA replication of several viruses, including the HCV replicon model. However, whether PI4P is required in an infectious HCV model remains unknown. Moreover, it is not established whether the host transport machinery is sequestered by the generation of PI4P during HCV infection. Here we found that PI4P was enriched in HCV replication complexes when Huh7.5.1 cells were infected with JFH1. HCV replication was inhibited upon overexpression of the PI4P phosphatase Sac1. The PI4P kinase PI4KIIIβ was also found to be required for HCV replication. Moreover, the vesicular transport proteins ARF1 and GBF1 colocalized with PI4KIIIβ and were both required for HCV replication. During authentic HCV infection, PI4P plays an integral role in virus replication.
Highlights
Hepatitis C virus (HCV), a positive-strand RNA virus of the family Flaviviridae, is one of the most prominent health threats in the world today, infecting more than 170 million persons globally [1]
Recent trials in patients with HCV genotype 1 infection combining the standard of care (SOC) with telaprevir or boceprevir have found that sustained virologic response (SVR) rates is 61–75% for telaprevir [5,6,7,8] or 67–75% for boceprevir [9,10,11]
We examined whether ARF1 colocalized with PI4KIIIb during HCV infection
Summary
Hepatitis C virus (HCV), a positive-strand RNA virus of the family Flaviviridae, is one of the most prominent health threats in the world today, infecting more than 170 million persons globally [1]. To explore the importance of PI4P in HCV replication, we overexpressed Sac and infected Huh7.5.1 cells with JFH1. When Huh7.5.1 cells were knocked down ARF1 by siRNA (Figure 5A and 5B) and infected with JFH1, HCV replication was reduced 10-fold as assessed by western blot (Figure 5A) and QPCR (Figure 5B). The normalized luciferase activities were divided by the normalized luciferase activity from mock treatment Both siRNAs could strongly reduce HCV replication (Figure 5C).These data indicate that ARF1 is required for HCV replication. GBF1 is required for HCV replication To examine whether GBF1 is critical for HCV replication, Huh7.5.1 cells were treated with the GBF1 inhibitor GCA or ARF1 inhibitor BFA, infected with JFH1, and assessed for HCV replication These data confirm that GBF1 is required for HCV replication
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