title: Genome-wide RNAi screen identifies SEC61A and VCP as conserved regulators of Sindbis virus entry.
abstract: Alphaviruses are a large class of insect-borne human pathogens and little is known about the host-factor requirements for infection. To identify such factors, we performed a genome-wide RNAi screen using model Drosophila cells and validated 94 genes that impacted infection of Sindbis virus (SINV), the prototypical alphavirus. We identified a conserved role for SEC61A and valosin-containing protein (VCP) in facilitating SINV entry in insects and mammals. SEC61A and VCP selectively regulate trafficking of the entry receptor NRAMP2, and loss or pharmacological inhibition of these proteins leads to altered NRAMP2 trafficking to lysosomal compartments and proteolytic digestion within lysosomes. NRAMP2 is the major iron transporter in cells, and loss of NRAMP2 attenuates intracellular iron transport. Thus, this study reveals genes and pathways involved in both infection and iron homeostasis that may serve as targets for antiviral therapeutics or for iron-imbalance disorders.
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Debasis
authors: D
authors: Panda
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Patrick P
authors: PP
authors: Rose
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Sheri L
authors: SL
authors: Hanna
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Beth
authors: B
authors: Gold
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Kaycie C
authors: KC
authors: Hopkins
authors: Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Physiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Randolph B
authors: RB
authors: Lyde
authors: Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Physiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors: Michael S
authors: MS
authors: Marks
authors: Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA; Penn Genome Frontiers Institute, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: cherrys@mail.med.upenn.edu.
authors: Sara
authors: S
authors: Cherry
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000330:Aedes / Q000235:genetics / Q000378:metabolism / Q000821:virology; D000818:Animals; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D004330:Drosophila / Q000235:genetics / Q000378:metabolism / Q000821:virology; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D049750:Genome, Insect; D006801:Humans; D007501:Iron / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D000069816:SEC Translocation Channels; D012845:Sindbis Virus / Q000472:pathogenicity*; D000074405:Valosin Containing Protein; D053586:Virus Internalization; D000099236:Solute Carrier Family 11, Member 2
chemical_list: D000251:Adenosine Triphosphatases; D027682:Cation Transport Proteins; D029721:Drosophila Proteins; D007501:Iron; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D000069816:SEC Translocation Channels; D000074405:Valosin Containing Protein; D000099236:Solute Carrier Family 11, Member 2; C585179:ter94 protein, Drosophila