Proteolytic Cleavage of VP1 in 'A' Particles of Coxsackievirus B3 Does Not Appear to Mediate Virus Uncoating by HeLa Cells
Department of Microbiology and Immunology, Hahnemann Medical College, Philadelphia, Pennsylvania 19102, U.S.A. A particles of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to A particles for...
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| Veröffentlicht in: | Journal of general virology 1981-08, Vol.55 (2), p.439-450 |
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| container_title | Journal of general virology |
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| creator | McGeady, Mary Lou Crowell, Richard L |
| description | Department of Microbiology and Immunology, Hahnemann Medical College, Philadelphia, Pennsylvania 19102, U.S.A.
A particles of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to A particles formed by elution from cellular receptors of HeLa cells. Electrophoretic analysis of [ 35 S]methionine-labelled A particles revealed that treatment of the particles with chymotrypsin resulted in the cleavage of VP1 and the formation of a cleavage product which migrated between VP2 and VP3. Analysis of the protease-treated material on sucrose gradients revealed a ribonuclease-sensitive particle which sedimented more slowly than an A particle. This particle apparently degraded to release the viral RNA, thereby providing an in vitro model for protease-mediated uncoating of A particles. The subviral particles of Coxsackievirus B3 were found to be immunoprecipitable with heterotypic Coxsackievirus group B antisera, thereby providing a method for the recovery of products produced in the cell early in infection. Infected cells which had been treated to remove unreacted virus were disrupted, and the lysates were reacted with heterotypic antisera. Analysis of the precipitated material revealed that no cleavage products were formed and no polypeptides were lost. Therefore, it appears that proteolysis is not involved in the uncoating of Coxsackievirus B3 in infected cells.
Keywords: Coxsackievirus B3, virus uncoating, proteolysis of A particles
Present Address: Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.
Received 26 November 1980;
accepted 13 March 1981. |
| doi_str_mv | 10.1099/0022-1317-55-2-439 |
| format | Article |
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A particles of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to A particles formed by elution from cellular receptors of HeLa cells. Electrophoretic analysis of [ 35 S]methionine-labelled A particles revealed that treatment of the particles with chymotrypsin resulted in the cleavage of VP1 and the formation of a cleavage product which migrated between VP2 and VP3. Analysis of the protease-treated material on sucrose gradients revealed a ribonuclease-sensitive particle which sedimented more slowly than an A particle. This particle apparently degraded to release the viral RNA, thereby providing an in vitro model for protease-mediated uncoating of A particles. The subviral particles of Coxsackievirus B3 were found to be immunoprecipitable with heterotypic Coxsackievirus group B antisera, thereby providing a method for the recovery of products produced in the cell early in infection. Infected cells which had been treated to remove unreacted virus were disrupted, and the lysates were reacted with heterotypic antisera. Analysis of the precipitated material revealed that no cleavage products were formed and no polypeptides were lost. Therefore, it appears that proteolysis is not involved in the uncoating of Coxsackievirus B3 in infected cells.
Keywords: Coxsackievirus B3, virus uncoating, proteolysis of A particles
Present Address: Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.
Received 26 November 1980;
accepted 13 March 1981.</description><identifier>ISSN: 0022-1317</identifier><identifier>EISSN: 1465-2099</identifier><identifier>DOI: 10.1099/0022-1317-55-2-439</identifier><identifier>PMID: 6270273</identifier><language>eng</language><publisher>England: Soc General Microbiol</publisher><subject>Capsid - analysis ; Capsid - metabolism ; Centrifugation, Density Gradient ; coxsackievirus B3 ; Enterovirus B, Human - growth & development ; Enterovirus B, Human - metabolism ; HeLa Cells ; Humans ; Peptide Hydrolases - metabolism ; RNA, Viral - metabolism ; Viral Proteins - metabolism ; Virion - metabolism</subject><ispartof>Journal of general virology, 1981-08, Vol.55 (2), p.439-450</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c405t-223963d98b13a48ba69244519cde58045537dff91bfbed34ec527df3cb1589a13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3732,3733,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6270273$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>McGeady, Mary Lou</creatorcontrib><creatorcontrib>Crowell, Richard L</creatorcontrib><title>Proteolytic Cleavage of VP1 in 'A' Particles of Coxsackievirus B3 Does Not Appear to Mediate Virus Uncoating by HeLa Cells</title><title>Journal of general virology</title><addtitle>J Gen Virol</addtitle><description>Department of Microbiology and Immunology, Hahnemann Medical College, Philadelphia, Pennsylvania 19102, U.S.A.
A particles of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to A particles formed by elution from cellular receptors of HeLa cells. Electrophoretic analysis of [ 35 S]methionine-labelled A particles revealed that treatment of the particles with chymotrypsin resulted in the cleavage of VP1 and the formation of a cleavage product which migrated between VP2 and VP3. Analysis of the protease-treated material on sucrose gradients revealed a ribonuclease-sensitive particle which sedimented more slowly than an A particle. This particle apparently degraded to release the viral RNA, thereby providing an in vitro model for protease-mediated uncoating of A particles. The subviral particles of Coxsackievirus B3 were found to be immunoprecipitable with heterotypic Coxsackievirus group B antisera, thereby providing a method for the recovery of products produced in the cell early in infection. Infected cells which had been treated to remove unreacted virus were disrupted, and the lysates were reacted with heterotypic antisera. Analysis of the precipitated material revealed that no cleavage products were formed and no polypeptides were lost. Therefore, it appears that proteolysis is not involved in the uncoating of Coxsackievirus B3 in infected cells.
Keywords: Coxsackievirus B3, virus uncoating, proteolysis of A particles
Present Address: Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.
Received 26 November 1980;
accepted 13 March 1981.</description><subject>Capsid - analysis</subject><subject>Capsid - metabolism</subject><subject>Centrifugation, Density Gradient</subject><subject>coxsackievirus B3</subject><subject>Enterovirus B, Human - growth & development</subject><subject>Enterovirus B, Human - metabolism</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Peptide Hydrolases - metabolism</subject><subject>RNA, Viral - metabolism</subject><subject>Viral Proteins - metabolism</subject><subject>Virion - metabolism</subject><issn>0022-1317</issn><issn>1465-2099</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1981</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUU1PGzEQtVARDSl_AKmST81piz-z62O6LQUp0BwgV8vrnQ1uN-tg7wLh19dpolx7Gs28N2_s9xC6pOQrJUpdEcJYRjnNMykzlgmuTtCIimlqEvwBjY6Ej-g8xt-EUCFkfobOpiwnLOcj9L4IvgffbntncdmCeTErwL7BywXFrsOT2QQvTEhoC3E3L_1bNPaPgxcXhoi_cfzdJ-Te93i22YAJuPf4DmpnesDLf5zHznrTu26Fqy2-gbnBJbRt_IROG9NGuDjUMXq8_vFQ3mTzXz9vy9k8s4LIPmOMqymvVVFRbkRRmali6RdU2RpkQYSUPK-bRtGqqaDmAqxkacBtRWWhDOVj9GWvuwn-eYDY67WLNr3AdOCHqHMuC0ZE_l8ilYIxxVQisj3RBh9jgEZvglubsNWU6F0yeme83hmvpdRMp2TS0ueD-lCtoT6uHKJI-GSPP7nV06sLoFfQrV06UTmvk9tHpb9YuZU7</recordid><startdate>198108</startdate><enddate>198108</enddate><creator>McGeady, Mary Lou</creator><creator>Crowell, Richard L</creator><general>Soc General Microbiol</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>198108</creationdate><title>Proteolytic Cleavage of VP1 in 'A' Particles of Coxsackievirus B3 Does Not Appear to Mediate Virus Uncoating by HeLa Cells</title><author>McGeady, Mary Lou ; Crowell, Richard L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c405t-223963d98b13a48ba69244519cde58045537dff91bfbed34ec527df3cb1589a13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1981</creationdate><topic>Capsid - analysis</topic><topic>Capsid - metabolism</topic><topic>Centrifugation, Density Gradient</topic><topic>coxsackievirus B3</topic><topic>Enterovirus B, Human - growth & development</topic><topic>Enterovirus B, Human - metabolism</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>Peptide Hydrolases - metabolism</topic><topic>RNA, Viral - metabolism</topic><topic>Viral Proteins - metabolism</topic><topic>Virion - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McGeady, Mary Lou</creatorcontrib><creatorcontrib>Crowell, Richard L</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of general virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>McGeady, Mary Lou</au><au>Crowell, Richard L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Proteolytic Cleavage of VP1 in 'A' Particles of Coxsackievirus B3 Does Not Appear to Mediate Virus Uncoating by HeLa Cells</atitle><jtitle>Journal of general virology</jtitle><addtitle>J Gen Virol</addtitle><date>1981-08</date><risdate>1981</risdate><volume>55</volume><issue>2</issue><spage>439</spage><epage>450</epage><pages>439-450</pages><issn>0022-1317</issn><eissn>1465-2099</eissn><abstract>Department of Microbiology and Immunology, Hahnemann Medical College, Philadelphia, Pennsylvania 19102, U.S.A.
A particles of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to A particles formed by elution from cellular receptors of HeLa cells. Electrophoretic analysis of [ 35 S]methionine-labelled A particles revealed that treatment of the particles with chymotrypsin resulted in the cleavage of VP1 and the formation of a cleavage product which migrated between VP2 and VP3. Analysis of the protease-treated material on sucrose gradients revealed a ribonuclease-sensitive particle which sedimented more slowly than an A particle. This particle apparently degraded to release the viral RNA, thereby providing an in vitro model for protease-mediated uncoating of A particles. The subviral particles of Coxsackievirus B3 were found to be immunoprecipitable with heterotypic Coxsackievirus group B antisera, thereby providing a method for the recovery of products produced in the cell early in infection. Infected cells which had been treated to remove unreacted virus were disrupted, and the lysates were reacted with heterotypic antisera. Analysis of the precipitated material revealed that no cleavage products were formed and no polypeptides were lost. Therefore, it appears that proteolysis is not involved in the uncoating of Coxsackievirus B3 in infected cells.
Keywords: Coxsackievirus B3, virus uncoating, proteolysis of A particles
Present Address: Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.
Received 26 November 1980;
accepted 13 March 1981.</abstract><cop>England</cop><pub>Soc General Microbiol</pub><pmid>6270273</pmid><doi>10.1099/0022-1317-55-2-439</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of general virology, 1981-08, Vol.55 (2), p.439-450 |
| issn | 0022-1317 1465-2099 |
| language | eng |
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| source | MEDLINE; Microbiology Society; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Capsid - analysis Capsid - metabolism Centrifugation, Density Gradient coxsackievirus B3 Enterovirus B, Human - growth & development Enterovirus B, Human - metabolism HeLa Cells Humans Peptide Hydrolases - metabolism RNA, Viral - metabolism Viral Proteins - metabolism Virion - metabolism |
| title | Proteolytic Cleavage of VP1 in 'A' Particles of Coxsackievirus B3 Does Not Appear to Mediate Virus Uncoating by HeLa Cells |
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