Abstract
We screened a siRNA library targeting human tyrosine kinases in Huh-7 cells and identified c-terminal Src kinase (Csk) as one of the kinases involved in dengue virus replication. Knock-down of Csk expression by siRNAs or inhibition of Csk by an inhibitor reduced dengue virus RNA levels but did not affect viral entry. Csk partially colocalized with viral replication compartments. Dengue infection was drastically reduced in cells lacking the three ubiquitous src family kinases, Src, Fyn and Yes. Csk knock-down in these cells failed to block dengue virus replication suggesting that the effect of Csk is via regulation of Src family kinases. Csk was found to be hyper-phosphorylated during dengue infection and inhibition of protein kinase A led to a block in Csk phosphorylation and dengue virus replication. Overexpression studies suggest an important role for the kinase and SH3 domains in this process. Our results identified a novel role for Csk as a host tyrosine kinase involved in dengue virus replication and provide further insights into the role of host factors in dengue replication.
Highlights
We screened a siRNA library targeting human tyrosine kinases in Huh-7 cells and identified c-terminal Src kinase (Csk) as one of the kinases involved in dengue virus replication
After excluding siRNAs that were affecting cell viability, we identified five Tyrosine kinases (TK) namely c-terminal src kinase (CSK), ephrin type B receptor-2 (EPHB2), insulin receptor (INSR), protein tyrosine kinase 6 (PTK6) and protein tyrosine kinase 9-like (PTK9L) that caused a ≥2-fold reduction in Dengue virus (DENV) titers in the culture supernatants without cytotoxicity
We have identified a new role for Csk in DENV RNA replication
Summary
We screened a siRNA library targeting human tyrosine kinases in Huh-7 cells and identified c-terminal Src kinase (Csk) as one of the kinases involved in dengue virus replication. Our results identified a novel role for Csk as a host tyrosine kinase involved in dengue virus replication and provide further insights into the role of host factors in dengue replication. SiRNA screens and inhibitor studies have identified receptor tyrosine kinases in Influenza virus entry and replication[11,12]. In this study we screened a siRNA library targeting human tyrosine kinases to identify TKs that are necessary for infection of DENV in Huh-7 cells. Our results identify a new role for Csk in flavivirus replication, which further enhances our understanding of viral replication and provides a new drug target for antiviral development
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