Sequences within Both the 5' UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus
This study mapped regions of genomic RNA (gRNA) important for packaging and propagation of mouse mammary tumor virus (MMTV). MMTV is a type B betaretrovirus which preassembles intracellularly, a phenomenon distinct from retroviruses that assemble the progeny virion at cell surface just before buddin...
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creator | Mustafa, Farah Al Amri, Dhuha Al Ali, Farah Al Sari, Noor Al Suwaidi, Sarah Jayanth, Preethi Philips, Pretty S Rizvi, Tahir A |
description | This study mapped regions of genomic RNA (gRNA) important for packaging and propagation of mouse mammary tumor virus (MMTV). MMTV is a type B betaretrovirus which preassembles intracellularly, a phenomenon distinct from retroviruses that assemble the progeny virion at cell surface just before budding such as the type C human and feline immunodeficiency viruses (HIV and FIV). Studies of FIV and Mason-Pfizer monkey virus (MPMV), a type D betaretrovirus with similar intracellular virion assembly processes as MMTV, have shown that the 5' untranslated region (5' UTR) and 5' end of gag constitute important packaging determinants for gRNA. Three series of MMTV transfer vectors containing incremental amounts of gag or 5' UTR sequences, or incremental amounts of 5' UTR in the presence of 400 nucleotides (nt) of gag were constructed to delineate the extent of 5' sequences that may be involved in MMTV gRNA packaging. Real time PCR measured the packaging efficiency of these vector RNAs into MMTV particles generated by co-transfection of MMTV Gag/Pol, vesicular stomatitis virus envelope glycoprotein (VSV-G Env), and individual transfer vectors into human 293T cells. Transfer vector RNA propagation was monitored by measuring transduction of target HeLaT4 cells following infection with viral particles containing a hygromycin resistance gene expression cassette on the packaged RNA. These results reveal that the 5' end of MMTV genome is critical for both gRNA packaging and propagation, unlike the recently delineated FIV and MPMV packaging determinants that have been shown to be of bipartite nature. |
doi_str_mv | 10.1371/journal.pone.0047088 |
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MMTV is a type B betaretrovirus which preassembles intracellularly, a phenomenon distinct from retroviruses that assemble the progeny virion at cell surface just before budding such as the type C human and feline immunodeficiency viruses (HIV and FIV). Studies of FIV and Mason-Pfizer monkey virus (MPMV), a type D betaretrovirus with similar intracellular virion assembly processes as MMTV, have shown that the 5' untranslated region (5' UTR) and 5' end of gag constitute important packaging determinants for gRNA. Three series of MMTV transfer vectors containing incremental amounts of gag or 5' UTR sequences, or incremental amounts of 5' UTR in the presence of 400 nucleotides (nt) of gag were constructed to delineate the extent of 5' sequences that may be involved in MMTV gRNA packaging. Real time PCR measured the packaging efficiency of these vector RNAs into MMTV particles generated by co-transfection of MMTV Gag/Pol, vesicular stomatitis virus envelope glycoprotein (VSV-G Env), and individual transfer vectors into human 293T cells. Transfer vector RNA propagation was monitored by measuring transduction of target HeLaT4 cells following infection with viral particles containing a hygromycin resistance gene expression cassette on the packaged RNA. 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Real time PCR measured the packaging efficiency of these vector RNAs into MMTV particles generated by co-transfection of MMTV Gag/Pol, vesicular stomatitis virus envelope glycoprotein (VSV-G Env), and individual transfer vectors into human 293T cells. Transfer vector RNA propagation was monitored by measuring transduction of target HeLaT4 cells following infection with viral particles containing a hygromycin resistance gene expression cassette on the packaged RNA. These results reveal that the 5' end of MMTV genome is critical for both gRNA packaging and propagation, unlike the recently delineated FIV and MPMV packaging determinants that have been shown to be of bipartite nature.</description><subject>Genetic aspects</subject><subject>Physiological aspects</subject><subject>RNA viruses</subject><subject>Virus replication</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNqFjztPwzAUhSMEEqXwDxjuBGJIcR52nLEgKJVatWpD1-rGcRKX1C5xwmPjpxMeQ5mY7tHRdz7pOs65RwZeEHnXG9PWGqvBzmg5ICSMCOcHTs-LA99lPgkO9_Kxc2LthhAacMZ6zsdSPrdSC2nhVTWl0nBjmhKaUgK9hMdkAagzGGEBw1rCooNVLTPITQ2zXaO2WMFYw0q9GJijeMJC6eJ7Mq_NDgtslNFgcpia1kqY4naL9Tsk7bYTrFTd2lPnKMfKyrPf23eS-7vk9sGdzEbj2-HELeKYuxF6PoaMiYhSGqW5TAllqYdcUPS7hnE_EhmXUc6ZkEKmLAwyluV5nHo0Fl7Qd65-tAVWcq20MLqRb02BrbXr8XKxHoYx9ztRyP9hZ6u_7MUeW0qsmtKaqv362-6Dn1llgHw</recordid><startdate>20121016</startdate><enddate>20121016</enddate><creator>Mustafa, Farah</creator><creator>Al Amri, Dhuha</creator><creator>Al Ali, Farah</creator><creator>Al Sari, Noor</creator><creator>Al Suwaidi, Sarah</creator><creator>Jayanth, Preethi</creator><creator>Philips, Pretty S</creator><creator>Rizvi, Tahir A</creator><general>Public Library of Science</general><scope>IOV</scope><scope>ISR</scope></search><sort><creationdate>20121016</creationdate><title>Sequences within Both the 5' UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus</title><author>Mustafa, Farah ; Al Amri, Dhuha ; Al Ali, Farah ; Al Sari, Noor ; Al Suwaidi, Sarah ; Jayanth, Preethi ; Philips, Pretty S ; Rizvi, Tahir A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g998-7a12a466c75557bfeb056b1a8c5a25576827cd8e7f86ceceb643d6dff9b159c13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Genetic aspects</topic><topic>Physiological aspects</topic><topic>RNA viruses</topic><topic>Virus replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mustafa, Farah</creatorcontrib><creatorcontrib>Al Amri, Dhuha</creatorcontrib><creatorcontrib>Al Ali, Farah</creatorcontrib><creatorcontrib>Al Sari, Noor</creatorcontrib><creatorcontrib>Al Suwaidi, Sarah</creatorcontrib><creatorcontrib>Jayanth, Preethi</creatorcontrib><creatorcontrib>Philips, Pretty S</creatorcontrib><creatorcontrib>Rizvi, Tahir A</creatorcontrib><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mustafa, Farah</au><au>Al Amri, Dhuha</au><au>Al Ali, Farah</au><au>Al Sari, Noor</au><au>Al Suwaidi, Sarah</au><au>Jayanth, Preethi</au><au>Philips, Pretty S</au><au>Rizvi, Tahir A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sequences within Both the 5' UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus</atitle><jtitle>PloS one</jtitle><date>2012-10-16</date><risdate>2012</risdate><volume>7</volume><issue>10</issue><spage>e47088</spage><pages>e47088-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>This study mapped regions of genomic RNA (gRNA) important for packaging and propagation of mouse mammary tumor virus (MMTV). 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Real time PCR measured the packaging efficiency of these vector RNAs into MMTV particles generated by co-transfection of MMTV Gag/Pol, vesicular stomatitis virus envelope glycoprotein (VSV-G Env), and individual transfer vectors into human 293T cells. Transfer vector RNA propagation was monitored by measuring transduction of target HeLaT4 cells following infection with viral particles containing a hygromycin resistance gene expression cassette on the packaged RNA. These results reveal that the 5' end of MMTV genome is critical for both gRNA packaging and propagation, unlike the recently delineated FIV and MPMV packaging determinants that have been shown to be of bipartite nature.</abstract><pub>Public Library of Science</pub><doi>10.1371/journal.pone.0047088</doi><tpages>e47088</tpages></addata></record> |
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source | DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
subjects | Genetic aspects Physiological aspects RNA viruses Virus replication |
title | Sequences within Both the 5' UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus |
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