In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene
The barley yellow dwarf virus (BYDV) coat protein gene is separated from an adjacent downstream open reading frame (ORF) by a single termination codon. Immunological analysis of this downstream "readthrough" region reveals multiple coat protein readthrough products. A full-length 72-kDa (P...
Gespeichert in:
| Veröffentlicht in: | Virology (New York, N.Y.) N.Y.), 1994-11, Vol.205 (1), p.290-299 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 299 |
|---|---|
| container_issue | 1 |
| container_start_page | 290 |
| container_title | Virology (New York, N.Y.) |
| container_volume | 205 |
| creator | Filichkin, S.A. Lister, R.M. McGrath, P.F. Young, M.J. |
| description | The barley yellow dwarf virus (BYDV) coat protein gene is separated from an adjacent downstream open reading frame (ORF) by a single termination codon. Immunological analysis of this downstream "readthrough" region reveals multiple coat protein readthrough products. A full-length 72-kDa (P72) coat protein-readthrough fusion product is detected in total lysates from infected cells. However, purified aphid transmissible virions contain only a 50-kDa (P50) coat protein-readthrough product. Virion-associated P50 lacks the C-terminal domain predicted by its ORF sequence. A separate 33-kDa polypeptide (P33) corresponding to the readthrough C-terminus domain is detected in the crude cellular membrane fraction. Site-directed and deletion mutational analysis demonstrate that the readthrough ORF is dispensable for BYDV replication and virion accumulation in protoplasts. In contrast, a mutant which results in a continuous fusion product of coat and readthrough sequences is not viable. Point mutations were used to map regions required for P50 and P72 synthesis. A model explaining the relationships between the three forms of the readthrough polypeptides is proposed. |
| doi_str_mv | 10.1006/viro.1994.1645 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_76846318</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S004268228471645X</els_id><sourcerecordid>76846318</sourcerecordid><originalsourceid>FETCH-LOGICAL-c370t-2127a19ab174032c36a4bb26c0d504fcd71065575948e3af04114b3bb99ded8c3</originalsourceid><addsrcrecordid>eNqFkM1r3DAQxUVpSDZpr70VdOrNm5EsS_YxX00CCYHQFgoFIUvjroLX2kj2pvvfV8suuYVcZni8N4_hR8gXBnMGIE_XPoY5axoxZ1JUH8iMQSMLKAX7SGYAghey5vyIHKf0BFkrBYfkUDWq4ryakT-3A_3l14Fe_VtFTMmHgZrB0ftpNGMWpqdneWySTzR0dFwgPTexxw39jX0fXujli4ldrohToo9o3LiIYfq7oNc44Cdy0Jk-4ef9PiE_v1_9uLgp7h6uby_O7gpbKhgLzrgyrDEtUwJKbktpRNtyacFVIDrrFANZVapqRI2l6UAwJtqybZvGoatteUK-7XpXMTxPmEa99Mnm_8yAYUpayVrIktXvBpmUUgCoHJzvgjaGlCJ2ehX90sSNZqC33PWWu95y11vu-eDrvnlql-he43vQ2a93PmYOa49RJ-txsOh8RDtqF_xb1f8BctWRRQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16664007</pqid></control><display><type>article</type><title>In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>ScienceDirect Journals (5 years ago - present)</source><creator>Filichkin, S.A. ; Lister, R.M. ; McGrath, P.F. ; Young, M.J.</creator><creatorcontrib>Filichkin, S.A. ; Lister, R.M. ; McGrath, P.F. ; Young, M.J.</creatorcontrib><description>The barley yellow dwarf virus (BYDV) coat protein gene is separated from an adjacent downstream open reading frame (ORF) by a single termination codon. Immunological analysis of this downstream "readthrough" region reveals multiple coat protein readthrough products. A full-length 72-kDa (P72) coat protein-readthrough fusion product is detected in total lysates from infected cells. However, purified aphid transmissible virions contain only a 50-kDa (P50) coat protein-readthrough product. Virion-associated P50 lacks the C-terminal domain predicted by its ORF sequence. A separate 33-kDa polypeptide (P33) corresponding to the readthrough C-terminus domain is detected in the crude cellular membrane fraction. Site-directed and deletion mutational analysis demonstrate that the readthrough ORF is dispensable for BYDV replication and virion accumulation in protoplasts. In contrast, a mutant which results in a continuous fusion product of coat and readthrough sequences is not viable. Point mutations were used to map regions required for P50 and P72 synthesis. A model explaining the relationships between the three forms of the readthrough polypeptides is proposed.</description><identifier>ISSN: 0042-6822</identifier><identifier>EISSN: 1096-0341</identifier><identifier>DOI: 10.1006/viro.1994.1645</identifier><identifier>PMID: 7975225</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>barley yellow dwarf virus ; Base Sequence ; Capsid - genetics ; Cells, Cultured ; DNA Mutational Analysis ; Escherichia coli - genetics ; Genes, Viral ; Luteovirus - genetics ; Molecular Sequence Data ; Oligodeoxyribonucleotides ; Point Mutation ; Recombinant Fusion Proteins - genetics ; Sequence Deletion ; Virion ; Virus Replication</subject><ispartof>Virology (New York, N.Y.), 1994-11, Vol.205 (1), p.290-299</ispartof><rights>1994 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c370t-2127a19ab174032c36a4bb26c0d504fcd71065575948e3af04114b3bb99ded8c3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/viro.1994.1645$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7975225$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Filichkin, S.A.</creatorcontrib><creatorcontrib>Lister, R.M.</creatorcontrib><creatorcontrib>McGrath, P.F.</creatorcontrib><creatorcontrib>Young, M.J.</creatorcontrib><title>In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene</title><title>Virology (New York, N.Y.)</title><addtitle>Virology</addtitle><description>The barley yellow dwarf virus (BYDV) coat protein gene is separated from an adjacent downstream open reading frame (ORF) by a single termination codon. Immunological analysis of this downstream "readthrough" region reveals multiple coat protein readthrough products. A full-length 72-kDa (P72) coat protein-readthrough fusion product is detected in total lysates from infected cells. However, purified aphid transmissible virions contain only a 50-kDa (P50) coat protein-readthrough product. Virion-associated P50 lacks the C-terminal domain predicted by its ORF sequence. A separate 33-kDa polypeptide (P33) corresponding to the readthrough C-terminus domain is detected in the crude cellular membrane fraction. Site-directed and deletion mutational analysis demonstrate that the readthrough ORF is dispensable for BYDV replication and virion accumulation in protoplasts. In contrast, a mutant which results in a continuous fusion product of coat and readthrough sequences is not viable. Point mutations were used to map regions required for P50 and P72 synthesis. A model explaining the relationships between the three forms of the readthrough polypeptides is proposed.</description><subject>barley yellow dwarf virus</subject><subject>Base Sequence</subject><subject>Capsid - genetics</subject><subject>Cells, Cultured</subject><subject>DNA Mutational Analysis</subject><subject>Escherichia coli - genetics</subject><subject>Genes, Viral</subject><subject>Luteovirus - genetics</subject><subject>Molecular Sequence Data</subject><subject>Oligodeoxyribonucleotides</subject><subject>Point Mutation</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Sequence Deletion</subject><subject>Virion</subject><subject>Virus Replication</subject><issn>0042-6822</issn><issn>1096-0341</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1r3DAQxUVpSDZpr70VdOrNm5EsS_YxX00CCYHQFgoFIUvjroLX2kj2pvvfV8suuYVcZni8N4_hR8gXBnMGIE_XPoY5axoxZ1JUH8iMQSMLKAX7SGYAghey5vyIHKf0BFkrBYfkUDWq4ryakT-3A_3l14Fe_VtFTMmHgZrB0ftpNGMWpqdneWySTzR0dFwgPTexxw39jX0fXujli4ldrohToo9o3LiIYfq7oNc44Cdy0Jk-4ef9PiE_v1_9uLgp7h6uby_O7gpbKhgLzrgyrDEtUwJKbktpRNtyacFVIDrrFANZVapqRI2l6UAwJtqybZvGoatteUK-7XpXMTxPmEa99Mnm_8yAYUpayVrIktXvBpmUUgCoHJzvgjaGlCJ2ehX90sSNZqC33PWWu95y11vu-eDrvnlql-he43vQ2a93PmYOa49RJ-txsOh8RDtqF_xb1f8BctWRRQ</recordid><startdate>19941115</startdate><enddate>19941115</enddate><creator>Filichkin, S.A.</creator><creator>Lister, R.M.</creator><creator>McGrath, P.F.</creator><creator>Young, M.J.</creator><general>Elsevier Inc</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>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19941115</creationdate><title>In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene</title><author>Filichkin, S.A. ; Lister, R.M. ; McGrath, P.F. ; Young, M.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c370t-2127a19ab174032c36a4bb26c0d504fcd71065575948e3af04114b3bb99ded8c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>barley yellow dwarf virus</topic><topic>Base Sequence</topic><topic>Capsid - genetics</topic><topic>Cells, Cultured</topic><topic>DNA Mutational Analysis</topic><topic>Escherichia coli - genetics</topic><topic>Genes, Viral</topic><topic>Luteovirus - genetics</topic><topic>Molecular Sequence Data</topic><topic>Oligodeoxyribonucleotides</topic><topic>Point Mutation</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Sequence Deletion</topic><topic>Virion</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Filichkin, S.A.</creatorcontrib><creatorcontrib>Lister, R.M.</creatorcontrib><creatorcontrib>McGrath, P.F.</creatorcontrib><creatorcontrib>Young, M.J.</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>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Virology (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Filichkin, S.A.</au><au>Lister, R.M.</au><au>McGrath, P.F.</au><au>Young, M.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene</atitle><jtitle>Virology (New York, N.Y.)</jtitle><addtitle>Virology</addtitle><date>1994-11-15</date><risdate>1994</risdate><volume>205</volume><issue>1</issue><spage>290</spage><epage>299</epage><pages>290-299</pages><issn>0042-6822</issn><eissn>1096-0341</eissn><abstract>The barley yellow dwarf virus (BYDV) coat protein gene is separated from an adjacent downstream open reading frame (ORF) by a single termination codon. Immunological analysis of this downstream "readthrough" region reveals multiple coat protein readthrough products. A full-length 72-kDa (P72) coat protein-readthrough fusion product is detected in total lysates from infected cells. However, purified aphid transmissible virions contain only a 50-kDa (P50) coat protein-readthrough product. Virion-associated P50 lacks the C-terminal domain predicted by its ORF sequence. A separate 33-kDa polypeptide (P33) corresponding to the readthrough C-terminus domain is detected in the crude cellular membrane fraction. Site-directed and deletion mutational analysis demonstrate that the readthrough ORF is dispensable for BYDV replication and virion accumulation in protoplasts. In contrast, a mutant which results in a continuous fusion product of coat and readthrough sequences is not viable. Point mutations were used to map regions required for P50 and P72 synthesis. A model explaining the relationships between the three forms of the readthrough polypeptides is proposed.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>7975225</pmid><doi>10.1006/viro.1994.1645</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0042-6822 |
| ispartof | Virology (New York, N.Y.), 1994-11, Vol.205 (1), p.290-299 |
| issn | 0042-6822 1096-0341 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_76846318 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; ScienceDirect Journals (5 years ago - present) |
| subjects | barley yellow dwarf virus Base Sequence Capsid - genetics Cells, Cultured DNA Mutational Analysis Escherichia coli - genetics Genes, Viral Luteovirus - genetics Molecular Sequence Data Oligodeoxyribonucleotides Point Mutation Recombinant Fusion Proteins - genetics Sequence Deletion Virion Virus Replication |
| title | In Vivo Expression and Mutational Analysis of the Barley Yellow Dwarf Virus Readthrough Gene |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-14T00%3A26%3A55IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=In%20Vivo%20Expression%20and%20Mutational%20Analysis%20of%20the%20Barley%20Yellow%20Dwarf%20Virus%20Readthrough%20Gene&rft.jtitle=Virology%20(New%20York,%20N.Y.)&rft.au=Filichkin,%20S.A.&rft.date=1994-11-15&rft.volume=205&rft.issue=1&rft.spage=290&rft.epage=299&rft.pages=290-299&rft.issn=0042-6822&rft.eissn=1096-0341&rft_id=info:doi/10.1006/viro.1994.1645&rft_dat=%3Cproquest_cross%3E76846318%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16664007&rft_id=info:pmid/7975225&rft_els_id=S004268228471645X&rfr_iscdi=true |