title: The non-structural 3 (NS3) protein of dengue virus type 2 interacts with human nuclear receptor binding protein and is associated with alterations in membrane structure.
abstract: Flaviviral infections produce a distinct array of virus-induced intracellular membrane alterations that are associated with the flaviviral replication machinery. Currently, it is still unknown which flaviviral protein(s) is/are responsible for this induction. Using yeast two-hybrid and co-immunoprecipitation analyses, we demonstrated that the NS3 protein of dengue virus type 2 interacted specifically with nuclear receptor binding protein (NRBP), a host cellular protein that influences trafficking between the endoplasmic reticulum (ER) and Golgi, and that interacts with Rac3, a member of the Rho-GTPase family. Co-expression of NS3 and NRBP in baby hamster kidney cells exhibited significant subcellular co-localization, and revealed the redistribution of NRBP from the cytoplasm to the perinuclear region. Furthermore, a set of membrane structures affiliated with the rough ER at the perinuclear region was induced in cells transfected with NS3. These structures are reminiscent of the virus-induced convoluted membranes previously observed in flavivirus-infected cells. This interaction between dengue viral and host cell proteins as well as the formation of the NS3-induced membrane structures suggest that NS3 may subvert the role of NRBP in ER-Golgi trafficking.
authors.affiliation: Programme in Infectious Diseases, Department of Microbiology, Faculty of Medicine, National University of Singapore, Kent Ridge, Singapore 117597, Singapore.
authors.forename: John J E
authors.initials: JJ
authors.lastname: Chua
authors.forename: Mary M L
authors.initials: MM
authors.lastname: Ng
authors.forename: Vincent T K
authors.initials: VT
authors.lastname: Chow
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003716:Dengue Virus / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D018870:Endoplasmic Reticulum, Rough / Q000473:pathology / Q000821:virology; D007425:Intracellular Membranes / Q000648:ultrastructure* / Q000821:virology; D018613:Microscopy, Confocal; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D021381:Protein Transport; D020365:RNA Helicases; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D012697:Serine Endopeptidases; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D011494:Protein Kinases; D020365:RNA Helicases; D018160:Receptors, Cytoplasmic and Nuclear; D012697:Serine Endopeptidases; D033921:Vesicular Transport Proteins; D017361:Viral Nonstructural Proteins; C410809:NRBP1 protein, human; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases