Human genome-wide RNAi screen reveals host factors required for enterovirus 71 replication

Abstract

Enterovirus 71 (EV71) is a neurotropic enterovirus without antivirals or vaccine, and its host-pathogen interactions remain poorly understood. Here we use a human genome-wide RNAi screen to identify 256 host factors involved in EV71 replication in human rhabdomyosarcoma cells. Enrichment analyses reveal overrepresentation in processes like mitotic cell cycle and transcriptional regulation. We have carried out orthogonal experiments to characterize the roles of selected factors involved in cell cycle regulation and endoplasmatic reticulum-associated degradation. We demonstrate nuclear egress of CDK6 in EV71 infected cells, and identify CDK6 and AURKB as resistance factors. NGLY1, which co-localizes with EV71 replication complexes at the endoplasmatic reticulum, supports EV71 replication. We confirm importance of these factors for EV71 replication in a human neuronal cell line and for coxsackievirus A16 infection. A small molecule inhibitor of NGLY1 reduces EV71 replication. This study provides a comprehensive map of EV71 host factors and reveals potential antiviral targets.

Enterovirus 71 protease 2Apro and 3Cpro differentially inhibit the cellular endoplasmic reticulum-associated degradation (ERAD) pathway via distinct mechanisms, and enterovirus 71 hijacks ERAD component p97 to promote its replication

“…A recent study demonstrated that p97 is essential to HCV replication and proposed that p97 is involved in the assembly of HCV replicase [91]. In addition, Wu et al identified p97 as a cellular factor involved in EV71 replication using a genome-wide RNAi screen [92]. The current study proposed that EV71 hijacks p97 from the disabled ERAD machinery, identified p97 as a host factor for EV71 replication, and demonstrated that p97 co-localized with the viral protein 2C on EV71-induced ROs.…”

An Integrated Systems Biology Approach Identifies the Proteasome as a Critical Host Machinery for ZIKV and DENV Replication

“…To validate whether host proteins enriched in the above biological processes and protein complexes were functionally involved in ZIKV infection, we carried out a siRNA knockdown assay similar to those used for other viruses [12,[34][35][36][37]. Specifically, 10,415 druggable target genes were individually knocked down and ZIKV NS1 protein level was measured using a high-throughput homogenous timeresolved fluorescence (HTRF) assay as a surrogate for viral load in ZIKV-infected HEK 293 cells.…”

2018 international meeting of the Global Virus Network

“…The incidence of human neurotropic enteroviruses causing HFMD in patients admitted to hospitals, including human enterovirus A71 (HEV-A71) infections associated with severe and even fatal neurological complications in young children, has steadily increased in Singapore during the past decade, causing a significant healthcare and economic burden (Teo and Chu, 2016). Using genomewide siRNA and miRNA screens, coupled with proteome profiling, his lab identified numerous host susceptibility and resistance factors affecting HEV-A71 infection (Leong et al, 2015;Wu et al, 2016). The host factors and pathways identified are subject of downstream studies at the Chu lab to identify their role in differential disease outcome and to develop novel intervention strategies.…”