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      Identification and characterization of a mouse mammary tumor virus protein uniquely expressed on the surface of BALB/cV mammary tumor cells

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      Virology
      Published by Elsevier Inc.

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          Abstract

          A unique subline of BALB/c mice, designated BALB/cV, exhibits an intermediate mammary tumor incidence (47%) and harbors a distinct milk-transmitted mouse mammary tumor virus (MMTV). The BALB/cV subline was used to study the molecular basis of potential virus-host interactions involving cell surface-expressed MMTV proteins. Cell surface iodination identified virus-specific proteins expressed on BALB/cv primary mammary tumor cells grown in culture. In contrast to (C3H)MMTV-producing cell lines which expressed MMTV gp52, BALB/cV tumor cells lacked gp52 and expressed instead a 68K, env-related protein. The 68K env protein was also detected on the surface of metabolically labeled BALB/cV tumor cells by an external immunoprecipitation technique. The expression of 68K env was restricted to mammary tissues of BALB/cV mice that also expressed other MMTV proteins. Biochemical analysis established that 68K env was not modified by N-linked glycosylation. 125I-labeled 68K env was rapidly released into the media of tumor cell cultures and was recovered both in the form of a soluble protein and in a 100,000 g pellet. The biologic function of this cell surface-expressed viral protein remains unknown.

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          Tunicamycin resistant glycosylation of coronavirus glycoprotein: demonstration of a novel type of viral glycoprotein.

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            Coronavirus glycoprotein E1, a new type of viral glycoprotein ☆

            The carbohydrate contents of coronavirus glycoproteins E1 and E2 have been analyzed. E2 has complex and mannose-rich-type oligosaccharide side-chains, which are attached by N-glycosidic linkages to the polypeptide. Glycosylation of E2 is initiated at the co-translational level, and it is inhibited by tunicamycin, 2-deoxy-glucose, and 2-deoxy-2-fluoro-glucose. Thus, E2 belongs to a glycoprotein type found in many other enveloped viruses. E1, in contrast, represents a different class of glycoprotein. The following observations indicate that its carbohydrate side-chains have 0-glycosidic linkage. (1) The constituent sugars of E1 are N-acetylglucosamine, N-acetylgalactosamine, galactose, and neuraminic acid; mannose and fucose are absent. (2) The side-chains can be removed by β-elimination. (3) Glycosylation of E1 is not sensitive to the compounds interfering with N-glycosylation. E1 is the first viral glycoprotein analyzed that contains only 0-glycosidic linkages. Coronaviruses are therefore a suitable model system to study biosynthesis and processing of this type of glycoprotein.
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              Tunicamycin inhibits glycosylation and multiplication of Sindbis and vesicular stomatitis viruses.

              Tunicamycin (TM), an antibiotic that inhibits the formation of N-acetylglucosamine-lipid intermediates, thereby preventing the glycosylation of newly synthesized glycoproteins, inhibits the growth of Sindbis virus and vesicular stomatitis virus in BHK cells. At 0.5 mug of TM per ml, the replication of both viruses is inhibited 99.9%. Noninfectious particles were not detected. All the viral proteins were synthesized in the presence of TM, but the glycoproteins were selectively altered in that they migrated faster than normal viral glycoproteins when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting defective glycosylation. Within 1 h after TM addition, [14C]glucosamine incorporation into glycoproteins was inhibited 20%, whereas [35S]methionine incorporation was unaffected. By 2 to 3 h after TM addition, glucosamine incorporation had fallen to 15% of control value, with methionine incorporation being 60% of normal. TM did not affect the growth of the nomenveloped encephalomyocarditis virus in BHK cells, demonstrating that TM is not a general inhibitor of protein synthesis. These data demonstrate that TM specifically inhibits the glycosylation of viral glycoproteins and that glycosylation may be essential for the normal assembly of enveloped viral particles.
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                Author and article information

                Journal
                Virology
                Virology
                Virology
                Published by Elsevier Inc.
                0042-6822
                1096-0341
                6 February 2004
                May 1985
                6 February 2004
                : 143
                : 1
                : 127-142
                Affiliations
                Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030, USA
                Author notes
                [1 ]Author to whom reprint requests should be addressed.
                Article
                0042-6822(85)90102-3
                10.1016/0042-6822(85)90102-3
                7130887
                2997998
                f4232050-35b1-4c10-89ec-391b7f57d92a
                Copyright © 1985 Published by Elsevier Inc.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 1 October 1984
                : 3 January 1985
                Categories
                Article

                Microbiology & Virology
                Microbiology & Virology

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