title: A physical interaction network of dengue virus and human proteins.
abstract: Dengue virus (DENV), an emerging mosquito-transmitted pathogen capable of causing severe disease in humans, interacts with host cell factors to create a more favorable environment for replication. However, few interactions between DENV and human proteins have been reported to date. To identify DENV-human protein interactions, we used high-throughput yeast two-hybrid assays to screen the 10 DENV proteins against a human liver activation domain library. From 45 DNA-binding domain clones containing either full-length viral genes or partially overlapping gene fragments, we identified 139 interactions between DENV and human proteins, the vast majority of which are novel. These interactions involved 105 human proteins, including six previously implicated in DENV infection and 45 linked to the replication of other viruses. Human proteins with functions related to the complement and coagulation cascade, the centrosome, and the cytoskeleton were enriched among the DENV interaction partners. To determine if the cellular proteins were required for DENV infection, we used small interfering RNAs to inhibit their expression. Six of 12 proteins targeted (CALR, DDX3X, ERC1, GOLGA2, TRIP11, and UBE2I) caused a significant decrease in the replication of a DENV replicon. We further showed that calreticulin colocalized with viral dsRNA and with the viral NS3 and NS5 proteins in DENV-infected cells, consistent with a direct role for calreticulin in DENV replication. Human proteins that interacted with DENV had significantly higher average degree and betweenness than expected by chance, which provides additional support for the hypothesis that viruses preferentially target cellular proteins that occupy central position in the human protein interaction network. This study provides a valuable starting point for additional investigations into the roles of human proteins in DENV infection.
authors.affiliation: Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Sudip
authors.initials: S
authors.lastname: Khadka
authors.forename: Abbey D
authors.initials: AD
authors.lastname: Vangeloff
authors.forename: Chaoying
authors.initials: C
authors.lastname: Zhang
authors.forename: Prasad
authors.initials: P
authors.lastname: Siddavatam
authors.forename: Nicholas S
authors.initials: NS
authors.lastname: Heaton
authors.forename: Ling
authors.initials: L
authors.lastname: Wang
authors.forename: Ranjan
authors.initials: R
authors.lastname: Sengupta
authors.forename: Sudhir
authors.initials: S
authors.lastname: Sahasrabudhe
authors.forename: Glenn
authors.initials: G
authors.lastname: Randall
authors.forename: Michael
authors.initials: M
authors.lastname: Gribskov
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Kuhn
authors.forename: Rushika
authors.initials: R
authors.lastname: Perera
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: LaCount
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism; D037282:Calreticulin / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000502:physiology*; D055785:Gene Knockdown Techniques; D017930:Genes, Reporter; D054884:Host-Pathogen Interactions; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D060066:Protein Interaction Maps; D021381:Protein Transport; D020365:RNA Helicases / Q000378:metabolism; D034622:RNA Interference; D012697:Serine Endopeptidases / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D014779:Virus Replication; D000098843:Ubiquitin-Conjugating Enzyme UBC9; D043583:Nucleoside-Triphosphatase; D000085242:Viral Proteases
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D001324:Autoantigens; D037282:Calreticulin; D003598:Cytoskeletal Proteins; D053487:DEAD-box RNA Helicases; D004279:DNA, Viral; D008156:Luciferases; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D020365:RNA Helicases; D012697:Serine Endopeptidases; D044763:Ubiquitin-Conjugating Enzymes; D017361:Viral Nonstructural Proteins; D000098843:Ubiquitin-Conjugating Enzyme UBC9; C468925:ERC1 protein, human; C097741:Golgin subfamily A member 2; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D000085242:Viral Proteases; C109331:TRIP11 protein, human; C119332:DDX3X protein, human