A 6K-deletion variant of salmonid alphavirus is non-viable but can be rescued through RNA recombination
Pancreas disease (PD) of Atlantic salmon is an emerging disease caused by Salmonid alphavirus (SAV) which mainly affects salmonid aquaculture in Western Europe. Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in vi...
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| Veröffentlicht in: | PloS one 2014-07, Vol.9 (7), p.e100184 |
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| creator | Guo, Tz-Chun Johansson, Daniel X Haugland, Øyvind Liljeström, Peter Evensen, Øystein |
| description | Pancreas disease (PD) of Atlantic salmon is an emerging disease caused by Salmonid alphavirus (SAV) which mainly affects salmonid aquaculture in Western Europe. Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in viral replication and infectivity remains undefined. The 6K protein of alphaviruses is a small and hydrophobic protein which is involved in membrane permeabilization, protein processing and virus budding. Because these common features are shared across many viral species, they have been named viroporins. In the present study, we applied reverse genetics to generate SAV3 6K-deleted (Δ6K) variant and investigate the role of 6K protein. Our findings show that the 6K-deletion variant of salmonid alphavirus is non-viable. Despite viral proteins of Δ6K variant are detected in the cytoplasm by immunostaining, they are not found on the cell surface. Further, analysis of viral proteins produced in Δ6K cDNA clone transfected cells using radioimmunoprecipitation (RIPA) and western blot showed a protein band of larger size than E2 of wild-type SAV3. When Δ6K cDNA was co-transfected with SAV3 helper cDNA encoding the whole structural genes including 6K, the infectivity was rescued. The development of CPE after co-transfection and resolved genome sequence of rescued virus confirmed full-length viral genome being generated through RNA recombination. The discovery of the important role of the 6K protein in virus production provides a new possibility for the development of antiviral intervention which is highly needed to control SAV infection in salmonids. |
| doi_str_mv | 10.1371/journal.pone.0100184 |
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Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in viral replication and infectivity remains undefined. The 6K protein of alphaviruses is a small and hydrophobic protein which is involved in membrane permeabilization, protein processing and virus budding. Because these common features are shared across many viral species, they have been named viroporins. In the present study, we applied reverse genetics to generate SAV3 6K-deleted (Δ6K) variant and investigate the role of 6K protein. Our findings show that the 6K-deletion variant of salmonid alphavirus is non-viable. Despite viral proteins of Δ6K variant are detected in the cytoplasm by immunostaining, they are not found on the cell surface. Further, analysis of viral proteins produced in Δ6K cDNA clone transfected cells using radioimmunoprecipitation (RIPA) and western blot showed a protein band of larger size than E2 of wild-type SAV3. When Δ6K cDNA was co-transfected with SAV3 helper cDNA encoding the whole structural genes including 6K, the infectivity was rescued. The development of CPE after co-transfection and resolved genome sequence of rescued virus confirmed full-length viral genome being generated through RNA recombination. The discovery of the important role of the 6K protein in virus production provides a new possibility for the development of antiviral intervention which is highly needed to control SAV infection in salmonids.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0100184</identifier><identifier>PMID: 25009976</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Alphavirus ; Alphavirus - drug effects ; Alphavirus - genetics ; Alphavirus - physiology ; Amino Acid Sequence ; Analysis ; Animals ; Antibodies - immunology ; Aquaculture ; Biology and Life Sciences ; Budding ; Cell Line ; Cell surface ; Clonal deletion ; Cytoplasm ; Cytoplasm - metabolism ; DNA, Complementary - genetics ; Fish ; Fishes ; Gene Deletion ; Genetics ; Genomes ; Genomics ; Health aspects ; Hydrophobicity ; Infectivity ; Interferon-alpha - pharmacology ; Medicine and Health Sciences ; Membrane proteins ; Mice ; Microbial Viability - genetics ; Molecular Sequence Data ; Molecular Weight ; Nucleotide sequence ; Oncorhynchus kisutch - virology ; Oncorhynchus mykiss ; Pancreas ; Proteins ; Recombination ; Recombination, Genetic ; Research and Analysis Methods ; Ribonucleic acid ; RNA ; RNA, Viral - genetics ; Salmo salar ; Salmon ; Salmonids ; Transfection ; Trout ; Viral Proteins - chemistry ; Viral Proteins - genetics ; Viral Proteins - immunology ; Viral Proteins - metabolism ; Virion - drug effects ; Virion - physiology ; Virus replication ; Viruses</subject><ispartof>PloS one, 2014-07, Vol.9 (7), p.e100184</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Guo et al. 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. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Guo et al 2014 Guo et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c730t-43362cdaabeacefe6693aa2b3c81351f43488417ce3a1d5c1cb3f2f864045cee3</citedby><cites>FETCH-LOGICAL-c730t-43362cdaabeacefe6693aa2b3c81351f43488417ce3a1d5c1cb3f2f864045cee3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4091863/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4091863/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,550,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79569,79570</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25009976$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttp://kipublications.ki.se/Default.aspx?queryparsed=id:129391375$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Guo, Tz-Chun</creatorcontrib><creatorcontrib>Johansson, Daniel X</creatorcontrib><creatorcontrib>Haugland, Øyvind</creatorcontrib><creatorcontrib>Liljeström, Peter</creatorcontrib><creatorcontrib>Evensen, Øystein</creatorcontrib><title>A 6K-deletion variant of salmonid alphavirus is non-viable but can be rescued through RNA recombination</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Pancreas disease (PD) of Atlantic salmon is an emerging disease caused by Salmonid alphavirus (SAV) which mainly affects salmonid aquaculture in Western Europe. Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in viral replication and infectivity remains undefined. The 6K protein of alphaviruses is a small and hydrophobic protein which is involved in membrane permeabilization, protein processing and virus budding. Because these common features are shared across many viral species, they have been named viroporins. In the present study, we applied reverse genetics to generate SAV3 6K-deleted (Δ6K) variant and investigate the role of 6K protein. Our findings show that the 6K-deletion variant of salmonid alphavirus is non-viable. Despite viral proteins of Δ6K variant are detected in the cytoplasm by immunostaining, they are not found on the cell surface. Further, analysis of viral proteins produced in Δ6K cDNA clone transfected cells using radioimmunoprecipitation (RIPA) and western blot showed a protein band of larger size than E2 of wild-type SAV3. When Δ6K cDNA was co-transfected with SAV3 helper cDNA encoding the whole structural genes including 6K, the infectivity was rescued. The development of CPE after co-transfection and resolved genome sequence of rescued virus confirmed full-length viral genome being generated through RNA recombination. The discovery of the important role of the 6K protein in virus production provides a new possibility for the development of antiviral intervention which is highly needed to control SAV infection in salmonids.</description><subject>Alphavirus</subject><subject>Alphavirus - drug effects</subject><subject>Alphavirus - genetics</subject><subject>Alphavirus - physiology</subject><subject>Amino Acid Sequence</subject><subject>Analysis</subject><subject>Animals</subject><subject>Antibodies - immunology</subject><subject>Aquaculture</subject><subject>Biology and Life Sciences</subject><subject>Budding</subject><subject>Cell Line</subject><subject>Cell surface</subject><subject>Clonal deletion</subject><subject>Cytoplasm</subject><subject>Cytoplasm - metabolism</subject><subject>DNA, Complementary - genetics</subject><subject>Fish</subject><subject>Fishes</subject><subject>Gene Deletion</subject><subject>Genetics</subject><subject>Genomes</subject><subject>Genomics</subject><subject>Health aspects</subject><subject>Hydrophobicity</subject><subject>Infectivity</subject><subject>Interferon-alpha - 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Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in viral replication and infectivity remains undefined. The 6K protein of alphaviruses is a small and hydrophobic protein which is involved in membrane permeabilization, protein processing and virus budding. Because these common features are shared across many viral species, they have been named viroporins. In the present study, we applied reverse genetics to generate SAV3 6K-deleted (Δ6K) variant and investigate the role of 6K protein. Our findings show that the 6K-deletion variant of salmonid alphavirus is non-viable. Despite viral proteins of Δ6K variant are detected in the cytoplasm by immunostaining, they are not found on the cell surface. Further, analysis of viral proteins produced in Δ6K cDNA clone transfected cells using radioimmunoprecipitation (RIPA) and western blot showed a protein band of larger size than E2 of wild-type SAV3. When Δ6K cDNA was co-transfected with SAV3 helper cDNA encoding the whole structural genes including 6K, the infectivity was rescued. The development of CPE after co-transfection and resolved genome sequence of rescued virus confirmed full-length viral genome being generated through RNA recombination. The discovery of the important role of the 6K protein in virus production provides a new possibility for the development of antiviral intervention which is highly needed to control SAV infection in salmonids.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25009976</pmid><doi>10.1371/journal.pone.0100184</doi><oa>free_for_read</oa></addata></record> |
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| ispartof | PloS one, 2014-07, Vol.9 (7), p.e100184 |
| issn | 1932-6203 1932-6203 |
| language | eng |
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| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; SWEPUB Freely available online; Free Full-Text Journals in Chemistry |
| subjects | Alphavirus Alphavirus - drug effects Alphavirus - genetics Alphavirus - physiology Amino Acid Sequence Analysis Animals Antibodies - immunology Aquaculture Biology and Life Sciences Budding Cell Line Cell surface Clonal deletion Cytoplasm Cytoplasm - metabolism DNA, Complementary - genetics Fish Fishes Gene Deletion Genetics Genomes Genomics Health aspects Hydrophobicity Infectivity Interferon-alpha - pharmacology Medicine and Health Sciences Membrane proteins Mice Microbial Viability - genetics Molecular Sequence Data Molecular Weight Nucleotide sequence Oncorhynchus kisutch - virology Oncorhynchus mykiss Pancreas Proteins Recombination Recombination, Genetic Research and Analysis Methods Ribonucleic acid RNA RNA, Viral - genetics Salmo salar Salmon Salmonids Transfection Trout Viral Proteins - chemistry Viral Proteins - genetics Viral Proteins - immunology Viral Proteins - metabolism Virion - drug effects Virion - physiology Virus replication Viruses |
| title | A 6K-deletion variant of salmonid alphavirus is non-viable but can be rescued through RNA recombination |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-13T13%3A47%3A10IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%206K-deletion%20variant%20of%20salmonid%20alphavirus%20is%20non-viable%20but%20can%20be%20rescued%20through%20RNA%20recombination&rft.jtitle=PloS%20one&rft.au=Guo,%20Tz-Chun&rft.date=2014-07-10&rft.volume=9&rft.issue=7&rft.spage=e100184&rft.pages=e100184-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0100184&rft_dat=%3Cgale_plos_%3EA418636010%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1544377435&rft_id=info:pmid/25009976&rft_galeid=A418636010&rft_doaj_id=oai_doaj_org_article_850a027f987c4795a54dd8e7a60fe8ae&rfr_iscdi=true |