Birdwell, Charles R. and Strauss, James H. (1974) Replication of Sindbis Virus IV. Electron Microscope Study of the Insertion of Viral Glycoproteins into the Surface of Infected Chick Cells. Journal of Virology, 14 (2). pp. 366-374. ISSN 0022-538X. http://resolver.caltech.edu/CaltechAUTHORS:BIRjvir74b
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The appearance of Sindbis virus-envelope glycoproteins in the surfaces of chicken embryo fibroblasts was studied by an indirect labeling technique. This technique involved treating infected cells sequentially with rabbit immunoglobulin G (IgG) specific for Sindbis virus followed by hemocyanin-conjugated goat (anti-rabbit IgG) IgG; surface replicas of these cells were then prepared and examined in the electron microscope. As early as 2 h after infection (and at least 1 h before mature virions were released), newly synthesized virus-envelope glycoproteins were detected at the cell surface. By 3 h after infection, cell surface membranes were extensively modified by the insertion of the Sindbis glycoproteins. When infected cells were prefixed with glutaraldehyde before labeling, the glycoproteins were distributed fairly evenly over the cell surface, although a slight clustering was observed on cells labeled early in infection. However, no evidence for large-scale clustering of virus glycoproteins corresponding to patches of budding virus was observed. Similar results were found with unfixed cells labeled at 4 C. However, when unfixed cells were labeled at 37 C, the glycoproteins were shown to be in discrete clusters, demonstrating that these glycoprotein antigens can diffuse laterally through the cell membrane at this temperature.
|Additional Information:||Copyright © 1974 American Society for Microbiology. Received for publication 15 April 1974. Jean-Paul Revel played an important role in the conduct of these experiments, pointing out the advantages of the surface replica technique and providing instruction in how to apply it. Ellen G. Strauss provided invaluable help in the preparation of the manuscript. Edith Lenches and Sharman Christoph provided competent technical assistance. This work was supported by grant GM 06965 from the U.S. Public Health Service and by grant GB 31763X from the National Science Foundation. These results are taken from the Ph.D. thesis of CRB [http://resolver.caltech.edu/CaltechETD:etd-09292005-083645], who was supported by U.S. Public Health Service Training grant GM 00086.|
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