RNA interference screen for human genes associated with West Nile virus infection.
West Nile virus (WNV), and related flaviviruses such as tick-borne encephalitis, Japanese encephalitis, yellow fever and dengue viruses, constitute a significant global human health problem. However, our understanding of the molecular interaction of such flaviviruses with mammalian host cells is limited. WNV encodes only 10 proteins, implying that it may use many cellular proteins for infection. WNV enters the cytoplasm through pH-dependent endocytosis, undergoes cycles of translation and replication, assembles progeny virions in association with endoplasmic reticulum, and exits along the secretory pathway. RNA interference (RNAi) presents a powerful forward genetics approach to dissect virus-host cell interactions. Here we report the identification of 305 host proteins that affect WNV infection, using a human-genome-wide RNAi screen. Functional clustering of the genes revealed a complex dependence of this virus on host cell physiology, requiring a wide variety of molecules and cellular pathways for successful infection. We further demonstrate a requirement for the ubiquitin ligase CBLL1 in WNV internalization, a post-entry role for the endoplasmic-reticulum-associated degradation pathway in viral infection, and the monocarboxylic acid transporter MCT4 as a viral replication resistance factor. By extending this study to dengue virus, we show that flaviviruses have both overlapping and unique interaction strategies with host cells. This study provides a comprehensive molecular portrait of WNV-human cell interactions that forms a model for understanding single plus-stranded RNA virus infection, and reveals potential antiviral targets.
Krishnan
Manoj N
MN
Section of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticutt 06520-8031, USA.
Ng
Aylwin
A
Sukumaran
Bindu
B
Gilfoy
Felicia D
FD
Uchil
Pradeep D
PD
Sultana
Hameeda
H
Brass
Abraham L
AL
Adametz
Rachel
R
Tsui
Melody
M
Qian
Feng
F
Montgomery
Ruth R
RR
Lev
Sima
S
Mason
Peter W
PW
Koski
Raymond A
RA
Elledge
Stephen J
SJ
Xavier
Ramnik J
RJ
Agaisse
Herve
H
Fikrig
Erol
E
D019295:Computational Biology; D003716:Dengue Virus / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D020869:Gene Expression Profiling; D015894:Genome, Human; D006678:HIV; D006367:HeLa Cells; D006801:Humans; D007109:Immunity / Q000235:genetics; D027501:Monocarboxylic Acid Transporters / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D009124:Muscle Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D034622:RNA Interference; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination / Q000235:genetics; D018116:Vesiculovirus; D014779:Virus Replication; D014901:West Nile Fever / Q000235:genetics* / Q000821:virology*; D014902:West Nile virus / Q000502:physiology*
D027501:Monocarboxylic Acid Transporters; D009124:Muscle Proteins; C406962:SLC16A4 protein, human; C471638:CBLL1 protein, human; D044767:Ubiquitin-Protein Ligases