scholarly journals Cell Entry of Borna Disease Virus Follows a Clathrin-Mediated Endocytosis Pathway That Requires Rab5 and Microtubules

2009 ◽  
Vol 83 (20) ◽  
pp. 10406-10416 ◽  
Author(s):  
Roberto Clemente ◽  
Juan C. de la Torre
Keyword(s):  
Disease Virus ◽  
Critical Role ◽  
Cell Entry ◽  
Surface Glycoprotein ◽  
Specific Cell ◽  
Critical Determinant ◽  
Borna Disease Virus ◽  
Glycoprotein G ◽  
The Central Nervous System ◽  
Borna Disease

ABSTRACT Borna disease virus (BDV), the prototypic member of the Bornaviridae family within the order Mononegavirales, exhibits high neurotropism and provides an important and unique experimental model system for studying virus-cell interactions within the central nervous system. BDV surface glycoprotein (G) plays a critical role in virus cell entry via receptor-mediated endocytosis, and therefore, G is a critical determinant of virus tissue and cell tropism. However, the specific cell pathways involved in BDV cell entry have not been determined. Here, we provide evidence that BDV uses a clathrin-mediated, caveola-independent cell entry pathway. We also show that BDV G-mediated fusion takes place at an optimal pH of 6.0 to 6.2, corresponding to an early-endosome compartment. Consistent with this finding, BDV cell entry was Rab5 dependent but Rab7 independent and exhibited rapid fusion kinetics. Our results also uncovered a key role for microtubules in BDV cell entry, whereas the integrity and dynamics of actin cytoskeleton were not required for efficient cell entry of BDV.

scholarly journals Identification of Host Factors Involved in Borna Disease Virus Cell Entry through a Small Interfering RNA Functional Genetic Screen

2010 ◽  
Vol 84 (7) ◽  
pp. 3562-3575 ◽  
Author(s):  
Roberto Clemente ◽  
Eugene Sisman ◽  
Pedro Aza-Blanc ◽  
Juan C. de la Torre
Keyword(s):  
Disease Virus ◽  
Small Interfering Rna ◽  
Cell Entry ◽  
Surface Glycoprotein ◽  
Cell Tropism ◽  
Borna Disease Virus ◽  
Virus Surface ◽  
Cellular Genes ◽  
Interfering Rna ◽  
Borna Disease

ABSTRACT 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 rVSVΔG*/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.

scholarly journals Generation and Characterization of a Recombinant Vesicular Stomatitis Virus Expressing the Glycoprotein of Borna Disease Virus

2007 ◽  
Vol 81 (11) ◽  
pp. 5527-5536 ◽  
Author(s):  
Mar Perez ◽  
Roberto Clemente ◽  
Clinton S. Robison ◽  
E. Jeetendra ◽  
Himangi R. Jayakar ◽  
...  
Keyword(s):  
Disease Virus ◽  
Vesicular Stomatitis Virus ◽  
Cell Entry ◽  
Surface Glycoprotein ◽  
Free Virus ◽  
Borna Disease Virus ◽  
Virus Family ◽  
Vesicular Stomatitis ◽  
Borna Disease

ABSTRACT Borna disease virus (BDV) is an enveloped virus with a nonsegmented negative-strand RNA genome whose organization is characteristic of mononegavirales. However, based on its unique genetics and biological features, BDV is considered to be the prototypic member of a new virus family, Bornaviridae, within the order Mononegavirales. BDV cell entry occurs via receptor-mediated endocytosis, a process initiated by the recognition of an as yet unidentified receptor at the cell surface by the BDV surface glycoprotein (G). The paucity of cell-free virus associated with BDV infection has hindered studies aimed at the elucidation of cellular receptors and detailed mechanisms involved in BDV cell entry. To overcome this problem, we generated and characterized a replication-competent recombinant vesicular stomatitis virus expressing BDV G (rVSVΔG*/BDVG). Cells infected with rVSVΔG*/BDVG produced high titers (107 PFU/ml) of cell-free virus progeny, but this virus exhibited a highly attenuated phenotype both in cell culture and in vivo. Attenuation of rVSVΔG*/BDVG was associated with a delayed kinetics of viral RNA replication and altered genome/N mRNA ratios compared to results for rVSVΔG*/VSVG. Likewise, incorporation of BDV G into virions appeared to be restricted despite its high levels of expression and efficient processing in rVSVΔG*/BDVG-infected cells. Notably, rVSVΔG*/BDVG recreated the cell tropism and entry pathway of bona fide BDV. Our results indicate that rVSVΔG*/BDVG represents a unique tool for the investigation of BDV G-mediated cell entry, as well as the roles of BDV G in host immune responses and pathogenesis associated with BDV infection.

scholarly journals Molecular Chaperone BiP Interacts with Borna Disease Virus Glycoprotein at the Cell Surface

2009 ◽  
Vol 83 (23) ◽  
pp. 12622-12625 ◽  
Author(s):  
Tomoyuki Honda ◽  
Masayuki Horie ◽  
Takuji Daito ◽  
Kazuyoshi Ikuta ◽  
Keizo Tomonaga
Keyword(s):  
Cell Surface ◽  
Disease Virus ◽  
Molecular Chaperone ◽  
Surface Glycoprotein ◽  
Target Cells ◽  
Borna Disease Virus ◽  
Virus Surface ◽  
Chaperone Protein ◽  
Glycoprotein G ◽  
Borna Disease

ABSTRACT Borna disease virus (BDV) is characterized by highly neurotropic infection. BDV enters its target cells using virus surface glycoprotein (G), but the cellular molecules mediating this process remain to be elucidated. We demonstrate here that the N-terminal product of G, GP1, interacts with the 78-kDa chaperone protein BiP. BiP was found at the surface of BDV-permissive cells, and anti-BiP antibody reduced BDV infection as well as GP1 binding to the cell surface. We also reveal that BiP localizes at the synapse of neurons. These results indicate that BiP may participate in the cell surface association of BDV.

scholarly journals Borna Disease Virus Requires Cholesterol in both Cellular Membrane and Viral Envelope for Efficient Cell Entry

2009 ◽  
Vol 83 (6) ◽  
pp. 2655-2662 ◽  
Author(s):  
Roberto Clemente ◽  
Aymeric de Parseval ◽  
Mar Perez ◽  
Juan C. de la Torre
Keyword(s):  
Disease Virus ◽  
Structural Integrity ◽  
Viral Envelope ◽  
Cell Entry ◽  
Surface Glycoprotein ◽  
Cholesterol Depletion ◽  
Borna Disease Virus ◽  
Virus Surface ◽  
And Function ◽  
Borna Disease

ABSTRACT Borna disease virus (BDV), the prototypic member of the family Bornaviridae within the order Mononegavirales, provides an important model for the investigation of viral persistence within the central nervous system (CNS) and of associated brain disorders. BDV is highly neurotropic and enters its target cell via receptor-mediated endocytosis, a process mediated by the virus surface glycoprotein (G), but the cellular factors and pathways determining BDV cell tropism within the CNS remain mostly unknown. Cholesterol has been shown to influence viral infections via its effects on different viral processes, including replication, budding, and cell entry. In this work, we show that cell entry, but not replication and gene expression, of BDV was drastically inhibited by depletion of cellular cholesterol levels. BDV G-mediated attachment to BDV-susceptible cells was cholesterol independent, but G localized to lipid rafts (LR) at the plasma membrane. LR structure and function critically depend on cholesterol, and hence, compromised structural integrity and function of LR caused by cholesterol depletion likely inhibited the initial stages of BDV cell internalization. Furthermore, we also show that viral-envelope cholesterol is required for BDV infectivity.

scholarly journals N-Terminal Domain of Borna Disease Virus G (p56) Protein Is Sufficient for Virus Receptor Recognition and Cell Entry

2001 ◽  
Vol 75 (15) ◽  
pp. 7078-7085 ◽  
Author(s):  
Mar Perez ◽  
Michiko Watanabe ◽  
Michael A. Whitt ◽  
Juan Carlos de la Torre
Keyword(s):  
Amino Acids ◽  
G Protein ◽  
Disease Virus ◽  
Fluorescent Protein ◽  
Virus Entry ◽  
Surface Glycoprotein ◽  
Terminal Part ◽  
Borna Disease Virus ◽  
Receptor Recognition ◽  
Borna Disease

ABSTRACT Borna disease virus (BDV) surface glycoprotein (GP) (p56) has a predicted molecular mass of 56 kDa. Due to extensive posttranslational glycosylation the protein migrates as a polypeptide of 84 kDa (gp84). The processing of gp84 by the cellular protease furin generates gp43, which corresponds to the C-terminal part of gp84. Both gp84 and gp43 have been implicated in viral entry involving receptor-mediated endocytosis and pH-dependent fusion. We have investigated the domains of BDV p56 involved in virus entry. For this, we used a pseudotype approach based on a recently developed recombinant vesicular stomatitis virus (VSV) in which the gene for green fluorescent protein was substituted for the VSV G protein gene (VSVΔG*). Complementation of VSVΔG* with BDV p56 resulted in infectious VSVΔG* pseudotypes that contained both BDV gp84 and gp43. BDV-VSV chimeric GPs that contained the N-terminal 244 amino acids of BDV p56 and amino acids 421 to 511 of VSV G protein were efficiently incorporated into VSVΔG* particles, and the resulting pseudotype virions were neutralized by BDV-specific antiserum. These findings indicate that the N-terminal part of BDV p56 is sufficient for receptor recognition and virus entry.

scholarly journals Borna Disease Virus Accelerates Inflammation and Disease Associated with Transgenic Expression of Interleukin-12 in the Central Nervous System

2002 ◽  
Vol 76 (23) ◽  
pp. 12223-12232 ◽  
Author(s):  
Susanna Freude ◽  
Jürgen Hausmann ◽  
Markus Hofer ◽  
Ngan Pham-Mitchell ◽  
Iain L. Campbell ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Transgenic Mice ◽  
Disease Virus ◽  
Biologically Active ◽  
Interleukin 12 ◽  
Wild Type ◽  
Borna Disease Virus ◽  
The Central Nervous System ◽  
Borna Disease

ABSTRACT Targeted expression of biologically active interleukin-12 (IL-12) in astrocytes of the central nervous system (CNS) results in spontaneous neuroimmunological disease of aged mice. Borna disease virus (BDV) can readily multiply in the mouse CNS but does not trigger disease in most strains. Here we show that a large percentage of IL-12 transgenic mice developed severe ataxia within 5 to 10 weeks after infection with BDV. By contrast, no disease developed in mock-infected IL-12 transgenic and wild-type mice until 4 months of age. Neurological symptoms were rare in infected wild-type animals, and if they occurred, these were milder and appeared later. Histological analyses showed that the cerebellum of infected IL-12 transgenic mice, which is the brain region with strongest transgene expression, contained large numbers of CD4+ and CD8+ T cells as well as lower numbers of B cells, whereas other parts of the CNS showed only mild infiltration by lymphocytes. The cerebellum of diseased mice further showed severe astrogliosis, calcifications and signs of neurodegeneration. BDV antigen and nucleic acids were present in lower amounts in the inflamed cerebellum of infected transgenic mice than in the noninflamed cerebellum of infected wild-type littermates, suggesting that IL-12 or IL-12-induced cytokines exhibited antiviral activity. We propose that BDV infection accelerates the frequency by which immune cells such as lymphocytes and NK cells enter the CNS and then respond to IL-12 present in the local milieu causing disease. Our results illustrate that infection of the CNS with a virus that is benign in certain hosts can be harmful in such normally disease-resistant hosts if the tissue is unfavorably preconditioned by proinflammatory cytokines.

scholarly journals Characterization of Borna disease virus p56 protein, a surface glycoprotein involved in virus entry.

1997 ◽  
Vol 71 (4) ◽  
pp. 3208-3218 ◽  
Author(s):  
D Gonzalez-Dunia ◽  
B Cubitt ◽  
F A Grasser ◽  
J C de la Torre
Keyword(s):  
Disease Virus ◽  
Virus Entry ◽  
Protein A ◽  
Surface Glycoprotein ◽  
Borna Disease Virus ◽  
Borna Disease

scholarly journals Borna Disease Virus Nucleoprotein Requires both Nuclear Localization and Export Activities for Viral Nucleocytoplasmic Shuttling

2001 ◽  
Vol 75 (7) ◽  
pp. 3404-3412 ◽  
Author(s):  
Takeshi Kobayashi ◽  
Wataru Kamitani ◽  
Guoqi Zhang ◽  
Makiko Watanabe ◽  
Keizo Tomonaga ◽  
...  
Keyword(s):  
Disease Virus ◽  
Nuclear Localization ◽  
Nuclear Export ◽  
Critical Role ◽  
Nuclear Export Signal ◽  
Nucleocytoplasmic Shuttling ◽  
Nuclear Targeting ◽  
Borna Disease Virus ◽  
Export Activity ◽  
Borna Disease

ABSTRACT Nuclear transport of viral nucleic acids is crucial to the life cycle of many viruses. Borna disease virus (BDV) belongs to the orderMononegavirales and replicates its RNA genome in the nucleus. Previous studies have suggested that BDV nucleoprotein (N) and phosphoprotein (P) have important functions in the nuclear import of the viral ribonucleoprotein (RNP) complexes via their nuclear targeting activity. Here, we showed that BDV N has cytoplasmic localization activity, which is mediated by a nuclear export signal (NES) within the sequence. Our analysis using deletion and substitution mutants of N revealed that NES of BDV N consists of a canonical leucine-rich motif and that the nuclear export activity of the protein is mediated through the chromosome region maintenance protein-dependent pathway. Interspecies heterokaryon assay indicated that BDV N shuttles between the nucleus and cytoplasm as a nucleocytoplasmic shuttling protein. Furthermore, interestingly, the NES region overlaps a binding site to the BDV P protein, and nuclear export of a 38-kDa form of BDV N is prevented by coexpression of P. These results suggested that BDV N has two contrary activities, nuclear localization and export activity, and plays a critical role in the nucleocytoplasmic transport of BDV RNP by interaction with other viral proteins.

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