title: The proteoglycan bamacan is a host cellular ligand of vaccinia virus neurovirulence factor N1L.
abstract: Neurovirulence is one of the pathological complications associated with vaccinia virus (VV) infection/vaccination. Although the viral N1L protein has been identified as the neurovirulence factor, none of the host N1L-interacting factors have been identified so far. In the present study, we identified N1L-interacting proteins by screening a human brain cDNA expression library with N1L as a bait protein in a yeast two-hybrid analysis. The analysis revealed that N1L interacts with human brain-originated cellular basement membrane-associated chondroitin sulfate proteoglycan (bamacan). The N1L-binding domain of bamacan was mapped to its C-terminal 227 amino acids. The N1L-bamacan interaction was further confirmed in both VV-infected and N1L-transfected mammalian cells. Following the confirmation of the protein interactions by coimmunoprecipitation experiments, confocal microscopic analysis revealed that N1L colocalizes with bamacan both in VV-infected B-SC-1 cells as well as in mice neuronal tissue. Furthermore, a human neural cell line, which expresses bamacan to moderately elevated levels relative to a non-neural cell line, supported enhanced viral growth. Overall, these studies clearly suggest that bamacan interacts with the VV-N1L and such interactions seem to play a positive role in promoting the viral growth and perhaps contribute to the virulence of VV in neural cells.
authors.affiliation: Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA. krishna.ketha391534@fda.hhs.gov
authors.forename: Ketha V K
authors.initials: KV
authors.lastname: Mohan
authors.forename: Cheryl X
authors.initials: CX
authors.lastname: Zhang
authors.forename: Chintamani D
authors.initials: CD
authors.lastname: Atreya
mesh_terms: D000818:Animals; D000833:Animals, Suckling; D001921:Brain / Q000378:metabolism / Q000821:virology*; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D008024:Ligands; D051379:Mice; D011485:Protein Binding; D014615:Vaccinia / Q000378:metabolism / Q000821:virology*; D014616:Vaccinia virus / Q000472:pathogenicity / Q000502:physiology*; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D011508:Chondroitin Sulfate Proteoglycans; D002868:Chromosomal Proteins, Non-Histone; D008024:Ligands; D014764:Viral Proteins; C492317:SMC3 protein, mouse; C464479:N1L protein, Vaccinia virus; C492318:SMC3 protein, human