Membrane topology and cellular dynamics of foot-and-mouth disease virus 3A protein
Foot-and-mouth disease virus non-structural protein 3A plays important roles in virus replication, virulence and host-range; nevertheless little is known on the interactions that this protein can establish with different cell components. In this work, we have performed in vivo dynamic studies from c...
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| description | Foot-and-mouth disease virus non-structural protein 3A plays important roles in virus replication, virulence and host-range; nevertheless little is known on the interactions that this protein can establish with different cell components. In this work, we have performed in vivo dynamic studies from cells transiently expressing the green fluorescent protein (GFP) fused to the complete 3A (GFP3A) and versions including different 3A mutations. The results revealed the presence of a mobile fraction of GFP3A, which was found increased in most of the mutants analyzed, and the location of 3A in a continuous compartment in the cytoplasm. A dual behavior was also observed for GFP3A upon cell fractionation, being the protein equally recovered from the cytosolic and membrane fractions, a ratio that was also observed when the insoluble fraction was further fractioned, even in the presence of detergent. Similar results were observed in the fractionation of GFP3ABBB, a 3A protein precursor required for initiating RNA replication. A nonintegral membrane protein topology of FMDV 3A was supported by the lack of glycosylation of versions of 3A in which each of the protein termini was fused to a glycosylation acceptor tag, as well as by their accessibility to degradation by proteases. According to this model 3A would interact with membranes through its central hydrophobic region exposing its N- and C- termini to the cytosol, where interactions between viral and cellular proteins required for virus replication are expected to occur. |
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In this work, we have performed in vivo dynamic studies from cells transiently expressing the green fluorescent protein (GFP) fused to the complete 3A (GFP3A) and versions including different 3A mutations. The results revealed the presence of a mobile fraction of GFP3A, which was found increased in most of the mutants analyzed, and the location of 3A in a continuous compartment in the cytoplasm. A dual behavior was also observed for GFP3A upon cell fractionation, being the protein equally recovered from the cytosolic and membrane fractions, a ratio that was also observed when the insoluble fraction was further fractioned, even in the presence of detergent. Similar results were observed in the fractionation of GFP3ABBB, a 3A protein precursor required for initiating RNA replication. A nonintegral membrane protein topology of FMDV 3A was supported by the lack of glycosylation of versions of 3A in which each of the protein termini was fused to a glycosylation acceptor tag, as well as by their accessibility to degradation by proteases. 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This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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According to this model 3A would interact with membranes through its central hydrophobic region exposing its N- and C- termini to the cytosol, where interactions between viral and cellular proteins required for virus replication are expected to occur.</description><subject>Amino acids</subject><subject>Animals</subject><subject>Biology and life sciences</subject><subject>Cell Line</subject><subject>Cell Membrane - metabolism</subject><subject>Cellular proteins</subject><subject>Cytoplasm</subject><subject>Cytosol</subject><subject>Endoplasmic reticulum</subject><subject>Fluorescence</subject><subject>Fluorescence Recovery After Photobleaching</subject><subject>Foot & mouth disease</subject><subject>Foot-and-mouth disease</subject><subject>Foot-and-Mouth Disease Virus - metabolism</subject><subject>Fractionation</subject><subject>Glycosylation</subject><subject>Green fluorescent protein</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Humans</subject><subject>Hydrophobicity</subject><subject>In vivo methods and tests</subject><subject>Membrane Microdomains - metabolism</subject><subject>Membrane proteins</subject><subject>Membranes</subject><subject>Microscopy</subject><subject>Mutant Proteins - chemistry</subject><subject>Mutant Proteins - metabolism</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Proteases</subject><subject>Protein 3a</subject><subject>Protein Binding</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Recovery (Medical)</subject><subject>Replication</subject><subject>Research and Analysis Methods</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Solubility</subject><subject>Topology</subject><subject>Transfection</subject><subject>Viral Proteins - metabolism</subject><subject>Virology</subject><subject>Virulence</subject><subject>Virus replication</subject><subject>Viruses</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNkluL3CAYhkNp6R7af1DaQKG0F5lqPCU3hWHpYWDLwvZwK2p0xiGJs2qWzr9f08kuk7IXxQtFn-_99PXNslcQLCBi8OPWDb4X7WLner0AEFBakSfZKaxRWdASoKdH65PsLIQtAARVlD7PTkpSMkIwPs2uv-tOetHrPLqda916n4u-yZVu26EVPm_2veisCrkzuXEuFum06NwQN3ljgxZB57fWDyFHy3znXdS2f5E9M6IN-uU0n2e_vnz-efGtuLz6urpYXhaK0TIWpqaVFkQKDJgoa1BLbbSoJaKNrFhVVphQYmolMcEGlKI0WFOIDSFGK0gbdJ69OejuWhf4ZEfgkFAGMEOMJWJ1IBontnznbSf8njth-d8N59dc-GhVqzmDDFEqmYQEYqmgEEw2SAmsAVGGyaT1aeo2yE43SvfRi3YmOj_p7Yav3S3HsEK4Gi_zfhLw7mbQIfLOhtHnZL4bxntXFNQUlyShb_9BH3_dRK1FeoDtjUt91SjKlxhSxlhVgUQtHqHSaHT615QdY9P-rODDrCAxUf-JazGEwFc_rv-fvfo9Z98dsRst2rgJrh2idX2Yg_gAKu9C8No8mAwBH6N_7wYfo8-n6Key18cf9FB0n3V0BzBx_o0</recordid><startdate>20141002</startdate><enddate>20141002</enddate><creator>González-Magaldi, Mónica</creator><creator>Martín-Acebes, Miguel A</creator><creator>Kremer, Leonor</creator><creator>Sobrino, Francisco</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>COVID</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20141002</creationdate><title>Membrane topology and cellular dynamics of foot-and-mouth disease virus 3A protein</title><author>González-Magaldi, Mónica ; Martín-Acebes, Miguel A ; Kremer, Leonor ; Sobrino, Francisco</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c762t-f968ea5ba407a2909befea9b36db878284565f9cb454f02a2f4e614f55fec16d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Amino acids</topic><topic>Animals</topic><topic>Biology and life sciences</topic><topic>Cell Line</topic><topic>Cell Membrane - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>González-Magaldi, Mónica</au><au>Martín-Acebes, Miguel A</au><au>Kremer, Leonor</au><au>Sobrino, Francisco</au><au>Li, Yi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Membrane topology and cellular dynamics of foot-and-mouth disease virus 3A protein</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-10-02</date><risdate>2014</risdate><volume>9</volume><issue>9</issue><spage>e106685</spage><epage>e106685</epage><pages>e106685-e106685</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Foot-and-mouth disease virus non-structural protein 3A plays important roles in virus replication, virulence and host-range; nevertheless little is known on the interactions that this protein can establish with different cell components. In this work, we have performed in vivo dynamic studies from cells transiently expressing the green fluorescent protein (GFP) fused to the complete 3A (GFP3A) and versions including different 3A mutations. The results revealed the presence of a mobile fraction of GFP3A, which was found increased in most of the mutants analyzed, and the location of 3A in a continuous compartment in the cytoplasm. A dual behavior was also observed for GFP3A upon cell fractionation, being the protein equally recovered from the cytosolic and membrane fractions, a ratio that was also observed when the insoluble fraction was further fractioned, even in the presence of detergent. Similar results were observed in the fractionation of GFP3ABBB, a 3A protein precursor required for initiating RNA replication. A nonintegral membrane protein topology of FMDV 3A was supported by the lack of glycosylation of versions of 3A in which each of the protein termini was fused to a glycosylation acceptor tag, as well as by their accessibility to degradation by proteases. According to this model 3A would interact with membranes through its central hydrophobic region exposing its N- and C- termini to the cytosol, where interactions between viral and cellular proteins required for virus replication are expected to occur.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25275544</pmid><doi>10.1371/journal.pone.0106685</doi><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; 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 | Amino acids Animals Biology and life sciences Cell Line Cell Membrane - metabolism Cellular proteins Cytoplasm Cytosol Endoplasmic reticulum Fluorescence Fluorescence Recovery After Photobleaching Foot & mouth disease Foot-and-mouth disease Foot-and-Mouth Disease Virus - metabolism Fractionation Glycosylation Green fluorescent protein Green Fluorescent Proteins - metabolism Humans Hydrophobicity In vivo methods and tests Membrane Microdomains - metabolism Membrane proteins Membranes Microscopy Mutant Proteins - chemistry Mutant Proteins - metabolism Mutants Mutation Proteases Protein 3a Protein Binding Proteins Recombinant Fusion Proteins - metabolism Recovery (Medical) Replication Research and Analysis Methods Ribonucleic acid RNA Solubility Topology Transfection Viral Proteins - metabolism Virology Virulence Virus replication Viruses |
| title | Membrane topology and cellular dynamics of foot-and-mouth disease virus 3A protein |
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