Lateral mobility of reconstituted Sendai virus envelope glycoproteins on human erythrocytes: correlation with cell-cell fusion

Biochemistry ◽  
1987 ◽  
Vol 26 (3) ◽  
pp. 812-819 ◽  
Author(s):  
Yoav I. Henis ◽  
Orit Gutman
Keyword(s):  
Cell Fusion ◽  
Sendai Virus ◽  
Human Erythrocytes ◽  
Envelope Glycoproteins ◽  
Lateral Mobility ◽  
Virus Envelope ◽  
Cell Cell

Studies of membrane fusion. VI. Mechanism of the membrane fusion and cell swelling stages of Sendai virus-mediated cell fusion

1980 ◽  
Vol 43 (1) ◽  
pp. 103-118
Author(s):  
S. Knutton
Keyword(s):  
Membrane Fusion ◽  
Cell Fusion ◽  
Sendai Virus ◽  
Freeze Fracture ◽  
Viral Envelope ◽  
Cell Swelling ◽  
Fusion Event ◽  
Virus Envelope ◽  
Membrane Rupture ◽  
Membrane Tubules

The membrane fusion and cell swelling stages of Sendai virus-mediated cell-cell fusion have been studied by thin-section and freeze-fracture electron microscopy. Sites of membrane fusion have been detected in human erythrocytes arrested at the membrane fusion stage of cell fusion and in virtually all cases a fused viral envelope or envelope components has been identified thus providing further direct evidence that cell-viral envelope-cell bridge formation is the membrane fusion event in Sendai virus-induced cell fusion. Radial expansion of a single virus bridge connecting 2 cells is sufficient to produce a fused cell. Membrane redistribution which occurs during this cell swelling stage of the fusion process is often accompanied by the formation of a system of membrane tubules in the plane of expansion of the virus bridge. The tubules originate from points of fusion between the bridging virus envelope and the erythrocyte membrane and also expand radially as cells swell. Ultimately membrane rupture occurs and the tubules appear to break down as small vesicles. When previously observed in cross-sectioned cells these membrane tubules were interpreted as sites of direct membrane fusion. The present study indicates that this interpretation is incorrect and shows that the tubules are generated subsequent to membrane fusion when 2 cells connected by a virus bridge are induced to swell. A mechanism to explain the formation of this system of membrane tubules is proposed.

The role of cell swelling and haemolysis in Sendai virus-induced cell fusion and in the diffusion of incorporated viral antigens

1980 ◽  
Vol 42 (1) ◽  
pp. 153-167
Author(s):  
S. Knutton ◽  
T. Bachi
Keyword(s):  
Cell Fusion ◽  
Erythrocyte Membrane ◽  
Sendai Virus ◽  
Viral Envelope ◽  
Cell Swelling ◽  
Haemolytic Activity ◽  
Erythrocyte Ghosts ◽  
Viral Antigens ◽  
Cell Cell

The role of the haemolytic activity of Sendai virus in cell-cell fusion has been examined in monolayers of human erythrocytes and erythrocyte ghosts fused with either haemolytic or non-haemolytic virus. Morphological observations indicate that cell swelling and haemolysis is a distinct event in cell-cell fusion irrespective of whether it is virally induced or, in the case of non-haemolytic virus, experimentally induced. Osmotic swelling appears to be the driving force by which cells which have established sites of membrane fusion expand such sites to form poly-erythrocytes. Immunofluorescent labelling of viral antigens incorporated into the erythrocyte membrane as a result of viral envelope-cell fusion indicates that diffusion of antigens in the plane of the membrane is restricted in intact erythrocytes and resealed erythrocyte ghosts but not in haemolysed erythrocytes or unsealed ghosts. A perturbation of the erythrocyte membrane resulting from osmotic lysis appears to form a prerequisite for the lateral diffusion of viral elements.

scholarly journals Functional Analysis of Hepatitis C Virus Envelope Proteins, Using a Cell-Cell Fusion Assay

2006 ◽  
Vol 80 (4) ◽  
pp. 1817-1825 ◽  
Author(s):  
Mariko Kobayashi ◽  
Michael C. Bennett ◽  
Theodore Bercot ◽  
Ila R. Singh
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Fusion ◽  
Viral Entry ◽  
Antiviral Agents ◽  
Cell Types ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
Host Cell Membrane ◽  
Cell Cell

ABSTRACT Hepatitis C virus (HCV) envelope proteins mediate the entry of virus into cells by binding to cellular receptors, resulting in fusion of the viral membrane with the host cell membrane and permitting the viral genome to enter the cytoplasm. We report the development of a robust and reproducible cell-cell fusion assay using envelope proteins from commonly occurring genotypes of HCV. The assay scored HCV envelope protein-mediated fusion by the production of fluorescent green syncytia and allowed us to elucidate many aspects of HCV fusion, including the pH of fusion, cell types that permit viral entry, and the conformation of envelope proteins essential for fusion. We found that fusion could be specifically inhibited by anti-HCV antibodies and by at least one peptide. We also generated a number of insertional mutations in the envelope proteins and tested nine of these using the fusion assay. We demonstrate that this fusion assay is a powerful tool for understanding the mechanism of HCV-mediated fusion, elucidating mutant function, and testing antiviral agents.

scholarly journals A 45,000-M(r) glycoprotein in the Sendai virus envelope triggers virus-cell fusion.

1997 ◽  
Vol 71 (9) ◽  
pp. 6398-6406 ◽  
Author(s):  
M Kumar ◽  
M Q Hassan ◽  
S K Tyagi ◽  
D P Sarkar
Keyword(s):  
Cell Fusion ◽  
Sendai Virus ◽  
Virus Envelope

scholarly journals "Ultramicroinjection" of macromolecules or small particles into animal cells. A new technique based on virus-induced cell fusion.

1975 ◽  
Vol 66 (2) ◽  
pp. 292-304 ◽  
Author(s):  
A Loyter ◽  
N Zakai ◽  
R G Kulka
Keyword(s):  
Cell Fusion ◽  
Sendai Virus ◽  
Human Erythrocytes ◽  
New Method ◽  
New Technique ◽  
Second Step ◽  
Small Particles ◽  
Animal Cells ◽  
Latex Spheres ◽  
A New Technique

A new method is described for the introduction of macromolecules and small particles into animal cells. The first step in this procedure is the trapping of particles in ghosts of human erythrocytes. This is achieved by the gradual hemolysis of erythrocytes in the presence of the particles to be trapped. The second step is the Sendai virus-induced fusion of the ghosts containing the particles with cells. By this method, ferritin and latex spheres (diameter 0.1 mum) have been "injected" into cells.

scholarly journals Rubella virus pseudotypes and a cell–cell fusion assay as tools for functional analysis of the rubella virus E2 and E1 envelope glycoproteins

2006 ◽  
Vol 87 (10) ◽  
pp. 3029-3037 ◽  
Author(s):  
Claudia Claus ◽  
Jörg Hofmann ◽  
Klaus Überla ◽  
U. G. Liebert
Keyword(s):  
Cell Fusion ◽  
Viral Entry ◽  
Rubella Virus ◽  
Lentiviral Vector ◽  
Envelope Glycoproteins ◽  
Immunodeficiency Virus ◽  
A Cell ◽  
Post Entry ◽  
In Cis ◽  
Cell Cell

The rubivirus Rubella virus contains the two envelope glycoproteins E2 and E1 as a heterodimeric spike complex embedded in its lipid envelope. The functions of both proteins, especially of E2, in the process of viral entry are still not entirely understood. In order to dissect E2 and E1 entry functions from post-entry steps, pseudotypes of lentiviral vectors based on Simian immunodeficiency virus were used. C-terminally modified E2 and E1 variants successfully pseudotyped lentiviral vector particles. This is the first report to show that not only E1, but also E2, is able to mediate infectious viral entry. Furthermore, a cell–cell fusion assay was used to further clarify membrane-fusion activities of E2 and E1 as one of the early steps of infection. It was demonstrated that the capsid protein, when coexpressed in cis, enhances the degree of E2- and E1-mediated cell–cell fusion.

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