Identification of host factors involved in borna disease virus cell entry through a small interfering RNA functional genetic screen.
Borna disease virus (BDV), the prototypic member of the Bornaviridae family, within the order Mononegavirales, is highly neurotropic and constitutes an important model system for the study of viral persistence in the central nervous system (CNS) and associated disorders. The virus surface glycoprotein (G) has been shown to direct BDV cell entry via receptor-mediated endocytosis, but the mechanisms governing cell tropism and propagation of BDV within the CNS are unknown. We developed a small interfering RNA (siRNA)-based screening to identify cellular genes and pathways that specifically contribute to BDV G-mediated cell entry. Our screen relied on silencing-mediated increased survival of cells infected with rVSVDeltaG*/BDVG, a cytolytic recombinant vesicular stomatitis virus expressing BDV G that mimics the cell tropism and entry pathway of bona fide BDV. We identified 24 cellular genes involved in BDV G-mediated cell entry. Identified genes are known to participate in a broad range of distinct cellular functions, revealing a complex process associated with BDV cell entry. The siRNA-based screening strategy we have developed should be applicable to identify cellular genes contributing to cell entry mediated by surface G proteins of other viruses.
Clemente
Roberto
R
Department of Immunology and Microbial Science, 10550 N. Torrey Pines Road, The Scripps Research Institute, La Jolla, CA 92037, USA.
Sisman
Eugene
E
Aza-Blanc
Pedro
P
de la Torre
Juan C
JC
D051722:ADAM Proteins / Q000502:physiology; D000072198:ADAM17 Protein; D001891:Borna disease virus / Q000502:physiology*; D002403:Cathepsins / Q000502:physiology; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000502:physiology; D056445:Cytoplasmic Dyneins / Q000502:physiology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D010447:Peptide Hydrolases / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics*; D053586:Virus Internalization
D034741:RNA, Small Interfering; D002403:Cathepsins; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; C117627:CTSV protein, human; D051722:ADAM Proteins; D000072198:ADAM17 Protein; D056445:Cytoplasmic Dyneins; C449283:DYNLRB2 protein, human