g protein-coupled receptor kinase 2 promotes flaviviridae entry and replicationg protein-coupled受体激酶2促进黄条目和复制.pdfVIP

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g protein-coupled receptor kinase 2 promotes flaviviridae entry and replicationg protein-coupled受体激酶2促进黄条目和复制.pdf

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g protein-coupled receptor kinase 2 promotes flaviviridae entry and replicationg protein-coupled受体激酶2促进黄条目和复制

G Protein-Coupled Receptor Kinase 2 Promotes Flaviviridae Entry and Replication Caroline Le Sommer1,2, Nicholas J. Barrows2,3,4, Shelton S. Bradrick1,2, James L. Pearson1,2,3, Mariano A. Garcia-Blanco1,2,5,6* 1 Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America, 2 Center for RNA Biology, Duke University Medical Center, Durham, North Carolina, United States of America, 3 Duke RNAi Screening Facility, Duke University Medical Center, Durham, North Carolina, United States of America, 4 Institute for Genome Sciences and Policy, Duke University, Durham, North Carolina, United States of America, 5 Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States of America, 6 Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore Abstract Flaviviruses cause a wide range of severe diseases ranging from encephalitis to hemorrhagic fever. Discovery of host factors that regulate the fate of flaviviruses in infected cells could provide insight into the molecular mechanisms of infection and therefore facilitate the development of anti-flaviviral drugs. We performed genome-scale siRNA screens to discover human host factors required for yellow fever virus (YFV) propagation. Using a 2 62 siRNA pool screening format and a duplicate of the screen, we identified a high confidence list of YFV host factors. To find commonalities between flaviviruses, these candidates were compared to host factors previously identified for West Nile virus (WNV) and dengue virus (DENV). This comparison highlighted a potential requirement for the G protein-coupled receptor kinase family, GRKs, for flaviviral infection. The YFV host candidate GRK2 (also known as ADRBK1) was validated both in siRNA-mediated knockdown HuH-7 cells and in GRK2/ 2 mouse e