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      Cholesterol is important for a post-adsorption step in the entry process of transmissible gastroenteritis virus

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          Abstract

          Cholesterol is a major constituent of detergent-resistant membrane microdomains (DRMs). We localized transmissible gastroenteritis virus (TGEV) spike (S) protein in DRMs in the viral envelope. Though S protein was not solubilized by cold non-ionic detergents, this behavior was unchanged when cholesterol was depleted from viral membrane by methyl-β-cyclodextrin (MβCD) and the protein did not comigrate with cellular DRM marker proteins in flotation analyses. Therefore, the S protein is not anchored in the viral membrane DRMs as they are known to occur in the plasma membrane. Cholesterol depletion from viral membrane may not affect the adsorption process as neither the sialic acid binding activity nor the binding to aminopeptidase N was reduced post-MβCD treatment. Reduced infectivity of cholesterol-depleted TGEV was observed only when the adsorption process occurred at 37 °C but not when the virus was applied at 4 °C. Cholesterol is important for a post-adsorption step, allowing membrane rearrangements that facilitate virus entry.

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          Most cited references35

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          Coronaviruses: structure and genome expression.

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            How viruses enter animal cells.

            Viruses replicate within living cells and use the cellular machinery for the synthesis of their genome and other components. To gain access, they have evolved a variety of elegant mechanisms to deliver their genes and accessory proteins into the host cell. Many animal viruses take advantage of endocytic pathways and rely on the cell to guide them through a complex entry and uncoating program. In the dialogue between the cell and the intruder, the cell provides critical cues that allow the virus to undergo molecular transformations that lead to successful internalization, intra-cellular transport, and uncoating.
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              Targeted recombination demonstrates that the spike gene of transmissible gastroenteritis coronavirus is a determinant of its enteric tropism and virulence.

              Targeted recombination within the S (spike) gene of transmissible gastroenteritis coronavirus (TGEV) was promoted by passage of helper respiratory virus isolates in cells transfected with a TGEV-derived defective minigenome carrying the S gene from an enteric isolate. The minigenome was efficiently replicated in trans and packaged by the helper virus, leading to the formation of true recombinant and pseudorecombinant viruses containing the S proteins of both enteric and respiratory TGEV strains in their envelopes. The recombinants acquired an enteric tropism, and their analysis showed that they were generated by homologous recombination that implied a double crossover in the S gene resulting in replacement of most of the respiratory, attenuated strain S gene (nucleotides 96 to 3700) by the S gene of the enteric, virulent isolate. The recombinant virus was virulent and rapidly evolved in swine testis cells by the introduction of point mutations and in-phase codon deletions in a domain of the S gene (nucleotides 217 to 665) previously implicated in the tropism of TGEV. The helper virus, with an original respiratory tropism, was also found in the enteric tract, probably because pseudorecombinant viruses carrying the spike proteins from the respiratory strain and the enteric virus in their envelopes were formed. These results demonstrated that a change in the tropism and virulence of TGEV can be engineered by sequence changes in the S gene.
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                Author and article information

                Contributors
                Journal
                Antiviral Res
                Antiviral Res
                Antiviral Research
                Elsevier B.V.
                0166-3542
                1872-9096
                14 October 2010
                December 2010
                14 October 2010
                : 88
                : 3
                : 311-316
                Affiliations
                [a ]College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, 150030 Harbin, China
                [b ]College of Life Sciences, Northeast Agricultural University, 59 Mucai Street, 150030 Harbin, China
                [c ]Institute for Virology, University of Veterinary Medicine Hannover, Bünteweg 17, D-30559 Hannover, Germany
                [d ]Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Campus Universitario de Cantoblanco, Darwin 3, 28049 Madrid, Spain
                Author notes
                [* ]Corresponding author. Tel.: +86 451 55190385; fax: +86 451 55103336. rxfemail@ 123456yahoo.com.cn renxf@ 123456neau.edu.cn
                Article
                S0166-3542(10)00744-8
                10.1016/j.antiviral.2010.10.002
                7114218
                20951168
                0018081b-dec9-4992-907f-f5dcc276e27d
                Copyright © 2010 Elsevier B.V. All rights reserved.

                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
                : 17 June 2010
                : 5 October 2010
                : 5 October 2010
                Categories
                Article

                Infectious disease & Microbiology
                lipid rafts,cholesterol,s protein,tgev,coronavirus
                Infectious disease & Microbiology
                lipid rafts, cholesterol, s protein, tgev, coronavirus

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