The Autographa californica Nuclear Polyhedrosis Virus ie2 Gene Encodes a Transcriptional Regulator
Identifieur interne : 001750 ( Istex/Corpus ); précédent : 001749; suivant : 001751The Autographa californica Nuclear Polyhedrosis Virus ie2 Gene Encodes a Transcriptional Regulator
Auteurs : Sunghan Yoo ; Linda A. GuarinoSource :
- Virology [ 0042-6822 ] ; 1994.
Abstract
Abstract: The baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) expresses several immediate early genes in infected cells. One of these, ie2 (formerly called IEN), was mapped and identified by its ability to augment IE1-mediated transactivation of the delayed early 39k gene. IE2 also stimulated the expression of reporter genes linked to the ie1 promoter. To test whether the increase in expression was due to increased mRNA, primer extension analyses were performed. Spodoptera frugiperda cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, in vitro transcription assays were performed. Nuclear extracts were prepared from S. frugiperda cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. Taken together, these results demonstrata that IE2 is a transcriptional regulator.
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DOI: 10.1006/viro.1994.1396
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Guarino</name><affiliation><mods:affiliation>Departments of Biochemistry & Biophysics, Entomology and The Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, Texas 77843-2475</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Virology</title><title level="j" type="abbrev">YVIRO</title><idno type="ISSN">0042-6822</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1994">1994</date><biblScope unit="volume">202</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="746">746</biblScope><biblScope unit="page" to="753">753</biblScope></imprint><idno type="ISSN">0042-6822</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0042-6822</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) expresses several immediate early genes in infected cells. One of these, ie2 (formerly called IEN), was mapped and identified by its ability to augment IE1-mediated transactivation of the delayed early 39k gene. IE2 also stimulated the expression of reporter genes linked to the ie1 promoter. To test whether the increase in expression was due to increased mRNA, primer extension analyses were performed. Spodoptera frugiperda cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, in vitro transcription assays were performed. Nuclear extracts were prepared from S. frugiperda cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. Taken together, these results demonstrata that IE2 is a transcriptional regulator.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Sunghan Yoo</name><affiliations><json:string>Departments of Biochemistry & Biophysics, Entomology and The Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, Texas 77843-2475</json:string></affiliations></json:item><json:item><name>Linda A. 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Spodoptera frugiperda cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, in vitro transcription assays were performed. Nuclear extracts were prepared from S. frugiperda cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. 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One of these, ie2 (formerly called IEN), was mapped and identified by its ability to augment IE1-mediated transactivation of the delayed early 39k gene. IE2 also stimulated the expression of reporter genes linked to the ie1 promoter. To test whether the increase in expression was due to increased mRNA, primer extension analyses were performed. Spodoptera frugiperda cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, in vitro transcription assays were performed. Nuclear extracts were prepared from S. frugiperda cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. Taken together, these results demonstrata that IE2 is a transcriptional regulator.</p></abstract></profileDesc><revisionDesc><change when="1994">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-WKKQHD16-M/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier converted-article found"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>YVIRO</jid><aid>71396</aid><ce:pii>S0042-6822(84)71396-1</ce:pii><ce:doi>10.1006/viro.1994.1396</ce:doi><ce:copyright type="full-transfer" year="1994">Academic Press</ce:copyright></item-info><head><ce:dochead><ce:textfn>Regular Article</ce:textfn></ce:dochead><ce:title>The <ce:italic>Autographa californica</ce:italic> Nuclear Polyhedrosis Virus <ce:italic>ie2</ce:italic> Gene Encodes a Transcriptional Regulator</ce:title><ce:author-group><ce:author><ce:given-name>Sunghan</ce:given-name><ce:surname>Yoo</ce:surname></ce:author><ce:author><ce:given-name>Linda A.</ce:given-name><ce:surname>Guarino</ce:surname></ce:author><ce:affiliation><ce:textfn>Departments of Biochemistry & Biophysics, Entomology and The Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, Texas 77843-2475</ce:textfn></ce:affiliation></ce:author-group><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>The baculovirus <ce:italic>Autographa californica</ce:italic> nuclear polyhedrosis virus (AcNPV) expresses several immediate early genes in infected cells. One of these, <ce:italic>ie2</ce:italic> (formerly called IEN), was mapped and identified by its ability to augment IE1-mediated transactivation of the delayed early <ce:italic>39k</ce:italic> gene. IE2 also stimulated the expression of reporter genes linked to the <ce:italic>ie1</ce:italic> promoter. To test whether the increase in expression was due to increased mRNA, primer extension analyses were performed. <ce:italic>Spodoptera frugiperda</ce:italic> cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, <ce:italic>in vitro</ce:italic> transcription assays were performed. Nuclear extracts were prepared from <ce:italic>S. frugiperda</ce:italic> cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. Taken together, these results demonstrata that IE2 is a transcriptional regulator.</ce:simple-para></ce:abstract-sec></ce:abstract></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>The Autographa californica Nuclear Polyhedrosis Virus ie2 Gene Encodes a Transcriptional Regulator</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>The</title></titleInfo><name type="personal"><namePart type="given">Sunghan</namePart><namePart type="family">Yoo</namePart><affiliation>Departments of Biochemistry & Biophysics, Entomology and The Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, Texas 77843-2475</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Linda A.</namePart><namePart type="family">Guarino</namePart><affiliation>Departments of Biochemistry & Biophysics, Entomology and The Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, Texas 77843-2475</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1994</dateIssued><copyrightDate encoding="w3cdtf">1994</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: The baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) expresses several immediate early genes in infected cells. One of these, ie2 (formerly called IEN), was mapped and identified by its ability to augment IE1-mediated transactivation of the delayed early 39k gene. IE2 also stimulated the expression of reporter genes linked to the ie1 promoter. To test whether the increase in expression was due to increased mRNA, primer extension analyses were performed. Spodoptera frugiperda cells were transfected with a plasmid encoding IE1 in the presence or the absence of a plasmid encoding IE2, and total cellular RNA was purified. Quantitative primer extension assays revealed that cells cotransfected with pIE1 and pIE2 contained higher levels of IE1 mRNA than cells transfected with pIE1 alone. To demonstrate directly that IE2 is a transcriptional regulator, in vitro transcription assays were performed. Nuclear extracts were prepared from S. frugiperda cells transfected with either pIE2 or pUC18. The extracts containing IE2 were more active than the control extracts in transcription of a C-free cassette linked to the IE1 promoter. Immunodepletion of IE2 from the transfected extracts reduced the transcription activity to that of the control extracts. Nuclear extracts prepared from IE2-transfected cells were also more active in transcription from the adenovirus major late promoter. 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