Phosphoproteomic-based kinase profiling early in influenza virus infection identifies GRK2 as antiviral drug target

Emilio Yángüez,Mirco Schmolke,Jonas Grossmann,Silke Stertz,Peter Gehrig,Ronald Dijkman,Umut Karakus,Soner Yildiz,Annika Hunziker,Maria Pamela Dobay,Simon Schading,Elizaveta Elshina

doi:10.1038/s41467-018-06119-y

Abstract

Although annual influenza epidemics affect around 10% of the global population, current treatment options are limited and development of new antivirals is needed. Here, using quantitative phosphoproteomics, we reveal the unique phosphoproteome dynamics that occur in the host cell within minutes of influenza A virus (IAV) infection. We uncover cellular kinases required for the observed signaling pattern and find that inhibition of selected candidates, such as the G protein-coupled receptor kinase 2 (GRK2), leads to decreased IAV replication. As GRK2 has emerged as drug target in heart disease, we focus on its role in IAV infection and show that it is required for viral uncoating. Replication of seasonal and pandemic IAVs is severely decreased by specific GRK2 inhibitors in primary human airway cultures and in mice. Our study reveals the IAV-induced changes to the cellular phosphoproteome and identifies GRK2 as crucial node of the kinase network that enables IAV replication.

Highlights

Annual influenza epidemics affect around 10% of the global population, current treatment options are limited and development of new antivirals is needed
While it was known before that this pathway becomes activated in response to IAV6,48, our study reveals its central role in early stages of influenza virus infection
We found G protein-coupled receptor kinase 2 (GRK2), ULK3, ROCK1 and p21 (RAC1) activated kinase 1 (PAK1) predicted to be involved in phosphorylation events identified at both time points analyzed

Summary

Introduction

Annual influenza epidemics affect around 10% of the global population, current treatment options are limited and development of new antivirals is needed. We uncover cellular kinases required for the observed signaling pattern and find that inhibition of selected candidates, such as the G protein-coupled receptor kinase 2 (GRK2), leads to decreased IAV replication. Binding of IAV particles, by interaction of the viral hemagglutinin (HA) to exposed sialylated proteins on epithelial cells[14], has been proposed to induce the formation of lipid raft-based signaling platforms, in which receptor tyrosine kinases (RTKs) such as the epidermal growth factor receptor (EGFR) or c-Met, are activated[6]. On the basis of this virus-induced phospho-signature, we are able to identify kinases, such as the G protein-coupled receptor kinase 2 (GRK2), that are activated during IAV entry and responsible for the observed signaling landscape. Our results establish GRK2 as a promising drug target for the generation of antivirals for influenza virus

Methods

Results

Conclusion