title: Use of a tandem affinity purification assay to detect interactions between West Nile and dengue viral proteins and proteins of the mosquito vector.
abstract: West Nile and dengue viruses are (re)emerging mosquito-borne flaviviruses that cause significant morbidity and mortality in man. The identification of mosquito proteins that associate with flaviviruses may provide novel targets to inhibit infection of the vector or block transmission to humans. Here, a tandem affinity purification (TAP) assay was used to identify 18 mosquito proteins that interact with dengue and West Nile capsid, envelope, NS2A or NS2B proteins. We further analyzed the interaction of mosquito cadherin with dengue and West Nile virus envelope protein using co-immunoprecipitation and immunofluorescence. Blocking the function of select mosquito factors, including actin, myosin, PI3-kinase and myosin light chain kinase, reduced both dengue and West Nile virus infection in mosquito cells. We show that the TAP method may be used in insect cells to accurately identify flaviviral-host protein interactions. Our data also provides several targets for interrupting flavivirus infection in mosquito vectors.
authors.affiliation: Section of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT, USA.
authors.forename: Tonya M
authors.initials: TM
authors.lastname: Colpitts
authors.forename: Jonathan
authors.initials: J
authors.lastname: Cox
authors.forename: Annie
authors.initials: A
authors.lastname: Nguyen
authors.forename: Fabiana
authors.initials: F
authors.lastname: Feitosa
authors.forename: Manoj N
authors.initials: MN
authors.lastname: Krishnan
authors.forename: Erol
authors.initials: E
authors.lastname: Fikrig
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D009033:Culicidae / Q000378:metabolism / Q000821:virology*; D003716:Dengue Virus / Q000378:metabolism*; D019476:Insect Proteins / Q000378:metabolism*; D007303:Insect Vectors / Q000378:metabolism / Q000821:virology*; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism*; D014902:West Nile virus / Q000378:metabolism*
chemical_list: D015820:Cadherins; D019476:Insect Proteins; D014764:Viral Proteins