Abstract
Type-I interferons (IFN) are key antiviral factors that induce several cellular proteins with antiviral activity. One such protein is Interferon Stimulated Gene 15 (ISG15). ISG15 is conjugated to proteins during ISGylation to confer antiviral activity. Apart from ISGylation, unconjugated ISG15 is also released from cells during immuno-stimulation and virus infection. The role of extracellular ISG15 during virus infection was unknown. We now show that extracellular ISG15 acts as a soluble antiviral factor to restrict virus infection in human epithelial cells via an IFN-independent mechanism. Additionally, we demonstrated that extracellular ISG15 induces expression of intracellular ISG15 and ISGylation in the absence of IFN. Mechanistic studies revealed that extracellular ISG15 interacts with cell surface integrin (α5β1 integrins) molecules via its RGD-like motif to activate the integrin-FAK (Focal Adhesion Kinase) pathway resulting in IFN-independent ISGylation. Thus, our studies have identified ISG15 protein as a new soluble factor that confers IFN-independent antiviral activity by inducing ISGylation. Additionally, we have unfolded an IFN-independent non-canonical ISGylation by extracellular ISG15 that operates via the integrin-FAK pathway.Type-I interferons (IFN) are key antiviral factors that induce several cellular proteins with antiviral activity. One such protein is Interferon Stimulated Gene 15 (ISG15). ISG15 is conjugated to proteins during ISGylation to confer antiviral activity. Apart from ISGylation, unconjugated ISG15 is also released from cells during immuno-stimulation and virus infection. The role of extracellular ISG15 during virus infection was unknown. We now show that extracellular ISG15 acts as a soluble antiviral factor to restrict virus infection in human epithelial cells via an IFN-independent mechanism. Additionally, we demonstrated that extracellular ISG15 induces expression of intracellular ISG15 and ISGylation in the absence of IFN. Mechanistic studies revealed that extracellular ISG15 interacts with cell surface integrin (α5β1 integrins) molecules via its RGD-like motif to activate the integrin-FAK (Focal Adhesion Kinase) pathway resulting in IFN-independent ISGylation. Thus, our studies have identified ISG15 protein as a new soluble factor that confers IFN-independent antiviral activity by inducing ISGylation. Additionally, we have unfolded an IFN-independent non-canonical ISGylation by extracellular ISG15 that operates via the integrin-FAK pathway.