scholarly journals Polarized expression of a chimeric protein in which the transmembrane and cytoplasmic domains of the influenza virus hemagglutinin have been replaced by those of the vesicular stomatitis virus G protein.

1986 ◽  
Vol 83 (24) ◽  
pp. 9318-9322 ◽  
Author(s):  
N. McQueen ◽  
D. P. Nayak ◽  
E. B. Stephens ◽  
R. W. Compans
Keyword(s):  
Influenza Virus ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Chimeric Protein ◽  
Cytoplasmic Domains ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis

Different properties of two isoforms of annexin XIII in MDCK cells

2000 ◽  
Vol 113 (14) ◽  
pp. 2607-2618 ◽  
Author(s):  
S. Lecat ◽  
P. Verkade ◽  
C. Thiele ◽  
K. Fiedler ◽  
K. Simons ◽  
...  
Keyword(s):  
Amino Acids ◽  
Influenza Virus ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Functional Data ◽  
Mdck Cells ◽  
Amino Terminal ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis ◽  
Amino Terminal Domain

Annexins form a family of proteins that are widely expressed and known to bind membranes in the presence of calcium. Two isoforms of the annexin XIII subfamily are expressed in epithelia. We previously reported that annexin XIIIb is apically localized in MDCK cells and that it is involved in raft-mediated delivery of apical proteins. We have now analyzed the properties of annexin XIIIa, which differs from annexin XIIIb by a deletion of 41 amino acids in the amino-terminal domain, and is distributed both apically and basolaterally. Annexin XIIIa binding to membranes is independent of calcium but requires its myristoyl amino-terminal modification, as observed with annexin XIIIb. Our biochemical and functional data show that annexin XIIIa behaves differently in the apical and in the basolateral compartments. Whereas annexin XIIIa apically can associate with rafts independently of calcium, the basolateral pool requires calcium for this. Annexin XIIIa, like annexin XIIIb, stimulates apical transport of influenza virus hemagglutinin but, in contrast, only annexin XIIIa inhibits basolateral transport of vesicular stomatitis virus G protein. Our results suggest that annexin XIIIa and XIIIb have specific roles in epithelial cells, and because of their structural similarities, these isoforms offer interesting tools for unravelling the functions of annexins.

scholarly journals Differential Targeting of Vesicular Stomatitis Virus G Protein and Influenza Virus Hemagglutinin Appears During Myogenesis of L6 Muscle Cells

1998 ◽  
Vol 140 (5) ◽  
pp. 1101-1111 ◽  
Author(s):  
P. Rahkila ◽  
V. Luukela ◽  
K. Väänänen ◽  
K. Metsikkö
Keyword(s):  
Influenza Virus ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Glucose Transporter ◽  
Myogenic Differentiation ◽  
Acquired Resistance ◽  
Brefeldin A ◽  
Myoblast Differentiation ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis

Exocytic organelles undergo profound reorganization during myoblast differentiation and fusion. Here, we analyzed whether glycoprotein processing and targeting changed during this process by using vesicular stomatitis virus (VSV) G protein and influenza virus hemagglutinin (HA) as models. After the induction of differentiation, the maturation and transport of the VSV G protein changed dramatically. Thus, only half of the G protein was processed and traveled through the Golgi, whereas the other half remained unprocessed. Experiments with the VSV tsO45 mutant indicated that the unprocessed form folded and trimerized normally and then exited the ER. It did not, however, travel through the Golgi since brefeldin A recalled it back to the ER. Influenza virus HA glycoprotein, on the contrary, acquired resistance to endoglycosidase H and insolubility in Triton X-100, indicating passage through the Golgi. Biochemical and morphological assays indicated that the HA appeared at the myotube surface. A major fraction of the Golgi-processed VSV G protein, however, did not appear at the myotube surface, but was found in intracellular vesicles that partially colocalized with the regulatable glucose transporter. Taken together, the results suggest that, during early myogenic differentiation, the VSV G protein was rerouted into developing, muscle-specific membrane compartments. Influenza virus HA, on the contrary, was targeted to the myotube surface.

scholarly journals The large external domain is sufficient for the correct sorting of secreted or chimeric influenza virus hemagglutinins in polarized monkey kidney cells.

1987 ◽  
Vol 104 (3) ◽  
pp. 769-782 ◽  
Author(s):  
M G Roth ◽  
D Gundersen ◽  
N Patil ◽  
E Rodriguez-Boulan
Keyword(s):  
Influenza Virus ◽  
Kidney Cells ◽  
Apical Surface ◽  
Collagen Gels ◽  
Cytoplasmic Domains ◽  
Gene Encoding ◽  
Ha Gene ◽  
Influenza Virus Hemagglutinin ◽  
External Domain ◽  
Vesicular Stomatitis

MA104.11 rhesus kidney cells express several characteristics of polarized epithelial cells, including the formation of "domes" on impermeable substrates, the establishment of a transmonolayer electrical resistance when grown on collagen gels, the polarized maturation of influenza and vesicular stomatitis viruses, and the expression of the glycoproteins of those viruses at a single surface domain. The polarized expression of the influenza virus hemagglutinin (HA) is maintained in MA104.11 cells infected with SV40-derived vectors carrying a cDNA gene for either the wild-type influenza virus HA, a truncated HA gene encoding a secreted form of HA (HAsec), or a chimeric gene encoding a hybrid protein with the external domain of the HA and the transmembrane and cytoplasmic domains of the vesicular stomatitis virus G protein (HAG). Thus, the recognition event separating glycoproteins, such as HA, destined for the apical surface from proteins, such as G, destined for the basolateral membranes involves features of the external domains of the proteins. The transmembrane and cytoplasmic domains of HA have no role in this process.

scholarly journals Display of Heterologous Proteins on gp64null Baculovirus Virions and Enhanced Budding Mediated by a Vesicular Stomatitis Virus G-Stem Construct

2007 ◽  
Vol 82 (3) ◽  
pp. 1368-1377 ◽  
Author(s):  
Jian Zhou ◽  
Gary W. Blissard
Keyword(s):  
Amino Acids ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Mammalian Cells ◽  
Heterologous Proteins ◽  
C Terminus ◽  
Influenza Virus Hemagglutinin ◽  
Severe Reduction ◽  
Vesicular Stomatitis ◽  
Ha Protein

ABSTRACT The Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) GP64 envelope glycoprotein is essential for virus entry and plays an important role in virion budding. An AcMNPV construct that contains a deletion of the gp64 gene is unable to propagate infection from cell to cell, and this defect results from both a severe reduction in the production of budded virions and the absence of GP64 on virions. In the current study, we examined GP64 proteins containing N- and C-terminal truncations of the ectodomain and identified a minimal construct capable of targeting the truncated GP64 to budded virions. The minimal budding and targeting construct of GP64 contained 38 amino acids from the mature N terminus of the GP64 ectodomain and 52 amino acids from the C terminus of GP64. Because the vesicular stomatitis virus (VSV) G protein was previously found to rescue infectivity of a gp64null AcMNPV, we also examined a small C-terminal construct of the VSV G protein. We found that a construct containing 91 amino acids from the C terminus of VSV G (termed G-stem) was capable of rescuing AcMNPV gp64null virion budding to wild-type (wt) or nearly wt levels. We also examined the display of chimeric proteins on the gp64null AcMNPV virion. By generating viruses that expressed chimeric influenza virus hemagglutinin (HA) proteins containing the GP64 targeting domain and coinfecting those viruses with a virus expressing the G-stem construct, we demonstrated enhanced display of the HA protein on gp64null AcMNPV budded virions. The combined use of gp64null virions, VSV G-stem-enhanced budding, and GP64 domains for targeting heterologous proteins to virions should be valuable for biotechnological applications ranging from targeted transduction of mammalian cells to vaccine production.

scholarly journals Attenuated Vesicular Stomatitis Viruses as Vaccine Vectors

1999 ◽  
Vol 73 (5) ◽  
pp. 3723-3732 ◽  
Author(s):  
Anjeanette Roberts ◽  
Linda Buonocore ◽  
Ryan Price ◽  
John Forman ◽  
John K. Rose
Keyword(s):  
Influenza Virus ◽  
Vesicular Stomatitis Virus ◽  
Neutralizing Antibodies ◽  
Intranasal Administration ◽  
Complete Protection ◽  
Lethal Challenge ◽  
Virus Challenge ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis ◽  
Ha Protein

ABSTRACT We showed previously that a single intranasal vaccination of mice with a recombinant vesicular stomatitis virus (VSV) expressing an influenza virus hemagglutinin (HA) protein provided complete protection from lethal challenge with influenza virus (A. Roberts, E. Kretzschmar, A. S. Perkins, J. Forman, R. Price, L. Buonocore, Y. Kawaoka, and J. K. Rose, J. Virol. 72:4704–4711, 1998). Because some pathogenesis was associated with the vector itself, in the present study we generated new VSV vectors expressing HA which are completely attenuated for pathogenesis in the mouse model. The first vector has a truncation of the cytoplasmic domain of the VSV G protein and expresses influenza virus HA (CT1-HA). This nonpathogenic vector provides complete protection from lethal influenza virus challenge after intranasal administration. A second vector with VSV G deleted and expressing HA (ΔG-HA) is also protective and nonpathogenic and has the advantage of not inducing neutralizing antibodies to the vector itself.

scholarly journals Vaccination with a Recombinant Vesicular Stomatitis Virus Expressing an Influenza Virus Hemagglutinin Provides Complete Protection from Influenza Virus Challenge

1998 ◽  
Vol 72 (6) ◽  
pp. 4704-4711 ◽  
Author(s):  
Anjeanette Roberts ◽  
Evelyne Kretzschmar ◽  
Archibald S. Perkins ◽  
John Forman ◽  
Ryan Price ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Vesicular Stomatitis Virus ◽  
Influenza A ◽  
Lethal Dose ◽  
Neutralizing Antibody ◽  
Virus Challenge ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis ◽  
High Level ◽  
Ha Protein

ABSTRACT Since the development of a system for generating vesicular stomatitis virus (VSV) from plasmid DNAs, our laboratory has reported the expression of several different glycoproteins from recombinant VSVs. In one of these studies, high-level expression of an influenza virus hemagglutinin (HA) from a recombinant VSV-HA and efficient incorporation of the HA protein into the virions was reported (E. Kretzschmar, L. Buonocore, M. J. Schnell, and J. K. Rose, J. Virol. 71:5982–5989, 1997). We report here that VSV-HA is an effective intranasal vaccine vector that raises high levels of neutralizing antibody to influenza virus and completely protects mice from bronchial pneumonia caused by challenge with a lethal dose of influenza A virus. Additionally, these recombinant VSVs are less pathogenic than wild-type VSV (serotype Indiana). This vector-associated pathogenicity was subsequently eliminated through introduction of specific attenuating deletions. These live attenuated recombinant VSVs have great potential as vaccine vectors.

scholarly journals A Viable Recombinant Rhabdovirus Lacking Its Glycoprotein Gene and Expressing Influenza Virus Hemagglutinin and Neuraminidase Is a Potent Influenza Vaccine

2014 ◽  
Vol 89 (5) ◽  
pp. 2820-2830 ◽  
Author(s):  
Alex B. Ryder ◽  
Linda Buonocore ◽  
Leatrice Vogel ◽  
Raffael Nachbagauer ◽  
Florian Krammer ◽  
...  
Keyword(s):  
Cell Culture ◽  
Influenza Virus ◽  
Influenza Vaccine ◽  
Vesicular Stomatitis Virus ◽  
Recombinant Virus ◽  
Vaccine Candidate ◽  
Influenza Vaccines ◽  
Glycoprotein Gene ◽  
Influenza Virus Hemagglutinin ◽  
Vesicular Stomatitis

ABSTRACTThe emergence of novel influenza viruses that cause devastating human disease is an ongoing threat and serves as an impetus for the continued development of novel approaches to influenza vaccines. Influenza vaccine development has traditionally focused on producing humoral and/or cell-mediated immunity, often against the viral surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). Here, we describe a new vaccine candidate that utilizes a replication-defective vesicular stomatitis virus (VSV) vector backbone that lacks the native G surface glycoprotein gene (VSVΔG). The expression of the H5 HA of an H5N1 highly pathogenic avian influenza virus (HPAIV), A/Vietnam/1203/04 (VN1203), and the NA of the mouse-adapted H1N1 influenza virus A/Puerto Rico/8/34 (PR8) in the VSVΔG vector restored the ability of the recombinant virus to replicate in cell culture, without the requirement for the addition of trypsin. We show here that this recombinant virus vaccine candidate was nonpathogenic in mice when given by either the intramuscular or intranasal route of immunization and that thein vivoreplication of VSVΔG-H5N1 is profoundly attenuated. This recombinant virus also provided protection against lethal H5N1 infection after a single dose. This novel approach to vaccination against HPAIVs may be widely applicable to other emerging strains of influenza virus.IMPORTANCEPreparation for a potentially catastrophic influenza pandemic requires novel influenza vaccines that are safe, can be produced and administered quickly, and are effective, both soon after administration and for a long duration. We have created a new influenza vaccine that utilizes an attenuated vesicular stomatitis virus (VSV) vector, to deliver and express influenza virus proteins against which vaccinated animals develop potent antibody responses. The influenza virus hemagglutinin and neuraminidase proteins, expressed on the surface of VSV particles, allowed this vaccine to grow in cell culture and induced a potent antibody response in mice that was effective against infection with a lethal influenza virus. The mice showed no adverse reactions to the vaccine, and they were protected against an otherwise lethal influenza infection after only 14 days postvaccination and after as many as 140 days postvaccination. The ability to rapidly produce this safe and effective vaccine in cell culture is additionally advantageous.

scholarly journals Caveolin-1 and -2 in the Exocytic Pathway of MDCK Cells

1998 ◽  
Vol 140 (4) ◽  
pp. 795-806 ◽  
Author(s):  
P. Scheiffele ◽  
P. Verkade ◽  
A.M. Fra ◽  
H. Virta ◽  
K. Simons ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Plasma Membrane ◽  
Vesicular Stomatitis Virus ◽  
Mdck Cells ◽  
Caveolin 1 ◽  
Apical Side ◽  
Influenza Virus Hemagglutinin ◽  
Basolateral Plasma Membrane ◽  
Vesicular Stomatitis ◽  
Exocytic Pathway

Abstract. We have studied the biosynthesis and transport of the endogenous caveolins in MDCK cells. We show that in addition to homooligomers of caveolin-1, heterooligomeric complexes of caveolin-1 and -2 are formed in the ER. The oligomers become larger, increasingly detergent insoluble, and phosphorylated on caveolin-2 during transport to the cell surface. In the TGN caveolin-1/-2 heterooligomers are sorted into basolateral vesicles, whereas larger caveolin-1 homooligomers are targeted to the apical side. Caveolin-1 is present on both the apical and basolateral plasma membrane, whereas caveolin-2 is enriched on the basolateral surface where caveolae are present. This suggests that caveolin-1 and -2 heterooligomers are involved in caveolar biogenesis in the basolateral plasma membrane. Anti–caveolin-1 antibodies inhibit the apical delivery of influenza virus hemagglutinin without affecting basolateral transport of vesicular stomatitis virus G protein. Thus, we suggest that caveolin-1 homooligomers play a role in apical transport.

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