Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA
Identifieur interne : 000979 ( Istex/Corpus ); précédent : 000978; suivant : 000980Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA
Auteurs : Béatrice Mestre ; Andrey Arzumanov ; Mohinder Singh ; Florence Boulmé ; Simon Litvak ; Michael J. GaitSource :
- BBA - Gene Structure and Expression [ 0167-4781 ] ; 1999.
English descriptors
- Teeft :
- Acta, Analogue, Antisense, Antisense oligonucleotides, Apical, Apical loop, Assay, Bbaexp, Biochimica, Biophysica, Biophysica acta, Bulge region, Chem, Control oligonucleotides, Hoechst, Human virus, Human virus type, Inhibition, Inhibitor, Karn, Lter, Lter binding, Lter binding assay, Lter binding interference assays, Lter retention, Mestre, Mobility shift analysis, Mobility shift assay, Native polyacrylamide, Neomycin, Nucleic acids, Oligodeoxyribonucleotides, Oligonucleotide, Oligonucleotide inhibition, Oligonucleotides, Oligoribonucleotide, Oligoribonucleotides, Oxford university press, Peptide, Peptide binding, Practical approach, Pyrimidine, Recognition sequence, Responsive region, Retarded band, Room temperature, Synthetic oligonucleotides, Transcription, Tris acetate.
Abstract
Abstract: The interaction of HIV-1 Tat protein with its recognition sequence, the trans-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-O-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-O-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.
Url:
DOI: 10.1016/S0167-4781(99)00019-6
Links to Exploration step
ISTEX:282A80C882C9652F58B797865503362587A61995Le document en format XML
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Gait</name><affiliation><mods:affiliation>E-mail: mgait@mrc-lmb.cam.ac.uk</mods:affiliation></affiliation><affiliation><mods:affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:282A80C882C9652F58B797865503362587A61995</idno><date when="1999" year="1999">1999</date><idno type="doi">10.1016/S0167-4781(99)00019-6</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000979</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000979</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA</title><author><name sortKey="Mestre, Beatrice" sort="Mestre, Beatrice" uniqKey="Mestre B" first="Béatrice" last="Mestre">Béatrice Mestre</name><affiliation><mods:affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</mods:affiliation></affiliation></author><author><name sortKey="Arzumanov, Andrey" sort="Arzumanov, Andrey" uniqKey="Arzumanov A" first="Andrey" last="Arzumanov">Andrey Arzumanov</name><affiliation><mods:affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</mods:affiliation></affiliation></author><author><name sortKey="Singh, Mohinder" sort="Singh, Mohinder" uniqKey="Singh M" first="Mohinder" last="Singh">Mohinder Singh</name><affiliation><mods:affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</mods:affiliation></affiliation></author><author><name sortKey="Boulme, Florence" sort="Boulme, Florence" uniqKey="Boulme F" first="Florence" last="Boulmé">Florence Boulmé</name><affiliation><mods:affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</mods:affiliation></affiliation></author><author><name sortKey="Litvak, Simon" sort="Litvak, Simon" uniqKey="Litvak S" first="Simon" last="Litvak">Simon Litvak</name><affiliation><mods:affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</mods:affiliation></affiliation></author><author><name sortKey="Gait, Michael J" sort="Gait, Michael J" uniqKey="Gait M" first="Michael J" last="Gait">Michael J. Gait</name><affiliation><mods:affiliation>E-mail: mgait@mrc-lmb.cam.ac.uk</mods:affiliation></affiliation><affiliation><mods:affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">BBA - Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="ISSN">0167-4781</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999">1999</date><biblScope unit="volume">1445</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="86">86</biblScope><biblScope unit="page" to="98">98</biblScope></imprint><idno type="ISSN">0167-4781</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4781</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acta</term><term>Analogue</term><term>Antisense</term><term>Antisense oligonucleotides</term><term>Apical</term><term>Apical loop</term><term>Assay</term><term>Bbaexp</term><term>Biochimica</term><term>Biophysica</term><term>Biophysica acta</term><term>Bulge region</term><term>Chem</term><term>Control oligonucleotides</term><term>Hoechst</term><term>Human virus</term><term>Human virus type</term><term>Inhibition</term><term>Inhibitor</term><term>Karn</term><term>Lter</term><term>Lter binding</term><term>Lter binding assay</term><term>Lter binding interference assays</term><term>Lter retention</term><term>Mestre</term><term>Mobility shift analysis</term><term>Mobility shift assay</term><term>Native polyacrylamide</term><term>Neomycin</term><term>Nucleic acids</term><term>Oligodeoxyribonucleotides</term><term>Oligonucleotide</term><term>Oligonucleotide inhibition</term><term>Oligonucleotides</term><term>Oligoribonucleotide</term><term>Oligoribonucleotides</term><term>Oxford university press</term><term>Peptide</term><term>Peptide binding</term><term>Practical approach</term><term>Pyrimidine</term><term>Recognition sequence</term><term>Responsive region</term><term>Retarded band</term><term>Room temperature</term><term>Synthetic oligonucleotides</term><term>Transcription</term><term>Tris acetate</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The interaction of HIV-1 Tat protein with its recognition sequence, the trans-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-O-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-O-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>oligonucleotides</json:string><json:string>oligonucleotide</json:string><json:string>antisense</json:string><json:string>lter</json:string><json:string>peptide</json:string><json:string>oligoribonucleotides</json:string><json:string>assay</json:string><json:string>acta</json:string><json:string>apical</json:string><json:string>bbaexp</json:string><json:string>biochimica</json:string><json:string>biophysica</json:string><json:string>mestre</json:string><json:string>biophysica acta</json:string><json:string>chem</json:string><json:string>apical loop</json:string><json:string>karn</json:string><json:string>oligoribonucleotide</json:string><json:string>lter binding assay</json:string><json:string>oligodeoxyribonucleotides</json:string><json:string>pyrimidine</json:string><json:string>hoechst</json:string><json:string>bulge region</json:string><json:string>mobility shift assay</json:string><json:string>neomycin</json:string><json:string>human virus</json:string><json:string>antisense oligonucleotides</json:string><json:string>nucleic acids</json:string><json:string>analogue</json:string><json:string>lter retention</json:string><json:string>lter binding</json:string><json:string>peptide binding</json:string><json:string>mobility shift analysis</json:string><json:string>oxford university press</json:string><json:string>human virus type</json:string><json:string>room temperature</json:string><json:string>synthetic oligonucleotides</json:string><json:string>native polyacrylamide</json:string><json:string>inhibitor</json:string><json:string>transcription</json:string><json:string>inhibition</json:string><json:string>tris acetate</json:string><json:string>retarded band</json:string><json:string>lter binding interference assays</json:string><json:string>oligonucleotide inhibition</json:string><json:string>responsive region</json:string><json:string>recognition sequence</json:string><json:string>control oligonucleotides</json:string><json:string>practical approach</json:string></teeft></keywords><author><json:item><name>Béatrice Mestre</name><affiliations><json:string>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</json:string></affiliations></json:item><json:item><name>Andrey Arzumanov</name><affiliations><json:string>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</json:string></affiliations></json:item><json:item><name>Mohinder Singh</name><affiliations><json:string>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</json:string></affiliations></json:item><json:item><name>Florence Boulmé</name><affiliations><json:string>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item><json:item><name>Simon Litvak</name><affiliations><json:string>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item><json:item><name>Michael J Gait</name><affiliations><json:string>E-mail: mgait@mrc-lmb.cam.ac.uk</json:string><json:string>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>HIV-1</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Tat protein</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>TAR RNA</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Oligonucleotide</value></json:item></subject><arkIstex>ark:/67375/6H6-7261QRW9-Z</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: The interaction of HIV-1 Tat protein with its recognition sequence, the trans-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-O-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-O-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.</abstract><qualityIndicators><score>9.1</score><pdfWordCount>6851</pdfWordCount><pdfCharCount>41674</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>13</pdfPageCount><pdfPageSize>596 x 794 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>175</abstractWordCount><abstractCharCount>1258</abstractCharCount><keywordCount>4</keywordCount></qualityIndicators><title>Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA</title><pmid><json:string>10209261</json:string></pmid><pii><json:string>S0167-4781(99)00019-6</json:string></pii><genre><json:string>research-article</json:string></genre><host><title>BBA - Gene Structure and Expression</title><language><json:string>unknown</json:string></language><publicationDate>1999</publicationDate><issn><json:string>0167-4781</json:string></issn><pii><json:string>S0167-4781(00)X0061-9</json:string></pii><volume>1445</volume><issue>1</issue><pages><first>86</first><last>98</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>2-4-99</json:string><json:string>1999</json:string></date><geogName></geogName><orgName><json:string>Cruachem</json:string><json:string>Institute for Molecular Biology</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Michael J. Gait</json:string><json:string>Serge Moreau</json:string><json:string>Terry Smith</json:string><json:string>Jan Fogg</json:string><json:string>David Owen</json:string><json:string>Glen Research</json:string><json:string>Bohrgasse</json:string><json:string>Richard Grenfell</json:string><json:string>Leo Saignat</json:string></persName><placeName><json:string>Bordeaux</json:string><json:string>Austria</json:string><json:string>Vienna</json:string><json:string>Puri</json:string><json:string>France</json:string><json:string>Saint Saens</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Hoechst 33258</json:string><json:string>[4]</json:string><json:string>[10,11]</json:string><json:string>[7,8]</json:string><json:string>[45]</json:string><json:string>[39,40]</json:string><json:string>[18]</json:string><json:string>[2,3]</json:string><json:string>[33]</json:string><json:string>[20,21]</json:string><json:string>[8]</json:string><json:string>[28]</json:string><json:string>[36,37]</json:string><json:string>[49,50]</json:string><json:string>[5,6]</json:string><json:string>B. Mestre et al.</json:string><json:string>[1]</json:string><json:string>[34,36]</json:string><json:string>[43]</json:string><json:string>[38]</json:string><json:string>[3]</json:string><json:string>[42]</json:string><json:string>[48]</json:string><json:string>[34,35]</json:string><json:string>[41]</json:string><json:string>[47]</json:string><json:string>[51]</json:string><json:string>[46]</json:string><json:string>[19]</json:string><json:string>[25,27]</json:string><json:string>Vickers et al.</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-7261QRW9-Z</json:string></ark><categories><wos></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - biochemistry & molecular biology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Genetics</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Biochemistry</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Biophysics</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Structural Biology</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences medicales</json:string><json:string>4 - radiotherapie. traitement instrumental. physiotherapie. reeducation. readaptation, orthophonie, crenotherapie. traitement dietetique et traitements divers (generalites)</json:string></inist></categories><publicationDate>1999</publicationDate><copyrightDate>1999</copyrightDate><doi><json:string>10.1016/S0167-4781(99)00019-6</json:string></doi><id>282A80C882C9652F58B797865503362587A61995</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a">Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>©1999 Elsevier Science B.V.</p></licence><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</p></availability><date>1999</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Fig. 1: (a) Regions of the HIV-1 Tat (BRU) protein and sequence of the Tat peptide (37–72). (b,c) Structures of the apical region of the HIV-1 TAR (BRU) and (MAL) stem-loops respectively as prepared synthetically, TAR BRU 39 and TAR MAL 38. The strain differences in the sequences of the loop and bulge region are boxed. For ease of comparison, the numbering scheme is harmonized between strains by considering nucleotide 25 to have been deleted in MAL (Δ25).</note><note type="content">Fig. 2: Competition filter binding interference assays showing inhibition of filter retention of TAR BRU 39 by Tat peptide (37–72) as a function of inhibitor concentration: (a) Hoechst 33258, neomycin B and a 16-mer oligonucleotide complementary to residues 22–37 (16 BRU (22–37)), (b) oligonucleotides 16 BRU (21–36), 16 BRU (22–37) and 16 BRU (23–38), (c) oligonucleotides 16 BRU (21–36), 15 BRU (22–36), 14 BRU (23–36) and 13 BRU (24–36), and (d) oligonucleotides 13 BRU (24–36), 11 BRU (26–36) and 6 BRU (31–36).</note><note type="content">Fig. 3: Competition filter binding interference assays showing inhibition of filter retention as a function of inhibitor concentration (a) for TAR BRU 39/Tat peptide (37–72) by oligonucleotides 15 BRU (21–35), 15 BRU mismatched and 15 BRU scrambled and (b) for TAR MAL 38/Tat protein by oligonucleotides 16 MAL (21–37), 16 MAL mismatched and 16 MAL scrambled.</note><note type="content">Fig. 4: Filter retention of 32P-labelled 16 BRU (23–38) oligonucleotide (a) as a function of concentration in the presence or absence of TAR BRU 39 and/or Tat peptide (37–72), and (b) as a function of increasing Tat peptide (37–72) concentration.</note><note type="content">Fig. 5: Competition filter binding interference assays showing inhibition of filter retention. (a) TAR BRU 39/Tat peptide (37–72) by oligonucleotides containing C5-propyne pyrimidines complementary to the loop (15 BRU L4), complementary to the stem (15 BRU S3), or both (15 BRU L4/S3) and unmodified oligonucleotide 15 BRU (21–35). The positions of C5-propyne incorporation are shown below underlined. (b) TAR MAL 38/Tat peptide (37–72) by all 2′-O-methyl oligoribonucleotides 16 MAL (21–37) OMe, 16 MAL mismatched OMe and 16 MAL scrambled (OMe) as well as unmodified 16 MAL (21–37) oligodeoxyribonucleotide.</note><note type="content">Fig. 6: Polyacrylamide gel mobility shift assay of binding of 2′-O-methyl oligoribonucleotides to a 274-residue 32P-labelled HIV-1 MAL transcript containing the full TAR RNA stem-loop. Lanes: 1, transcript alone; 2–6, 16 MAL OMe scrambled; 7–11, 16 MAL OMe mismatched; 12–16, 16 MAL OMe (21–37). The sequences of the oligonucleotides are shown in the lower part of Fig. 5.</note><note type="content">Fig. 7: Polyacrylamide gel mobility shift competition assay. Lanes: 1, 32P-labelled TAR MAL 38 alone; 2, TAR MAL 38+Tat peptide (37–72); 3, TAR MAL 38+16 MAL OMe (21–37); 4–6, TAR MAL 38, Tat peptide and increasing concentrations of 16 MAL OMe; 7, TAR MAL 38+mismatched OMe oligonucleotide; 8–10, TAR MAL 38, Tat peptide+increasing concentrations of mismatched OMe oligonucleotide; 11, TAR MAL 38+ scrambled OMe oligonucleotide; 12–14, TAR MAL 38, Tat peptide+increasing concentrations of scrambled OMe oligonucleotide.</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA</title><author xml:id="author-0000"><persName><forename type="first">Béatrice</forename><surname>Mestre</surname></persName><note type="biography">Present address: Laboratoire de Biophysique Moléculaire, INSERM U-386, Université Victor Ségalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux, France.</note><affiliation>Present address: Laboratoire de Biophysique Moléculaire, INSERM U-386, Université Victor Ségalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux, France.</affiliation><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Andrey</forename><surname>Arzumanov</surname></persName><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Mohinder</forename><surname>Singh</surname></persName><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Florence</forename><surname>Boulmé</surname></persName><note type="biography">Present address: Institute for Molecular Biology, Dr. Bohrgasse 9, 1030 Vienna, Austria.</note><affiliation>Present address: Institute for Molecular Biology, Dr. Bohrgasse 9, 1030 Vienna, Austria.</affiliation><affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Simon</forename><surname>Litvak</surname></persName><affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</affiliation></author><author xml:id="author-0005"><persName><forename type="first">Michael J</forename><surname>Gait</surname></persName><email>mgait@mrc-lmb.cam.ac.uk</email><note type="correspondence"><p>Corresponding author. Fax: +44 (1223) 402070</p></note><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation></author><idno type="istex">282A80C882C9652F58B797865503362587A61995</idno><idno type="ark">ark:/67375/6H6-7261QRW9-Z</idno><idno type="DOI">10.1016/S0167-4781(99)00019-6</idno><idno type="PII">S0167-4781(99)00019-6</idno></analytic><monogr><title level="j">BBA - Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="pISSN">0167-4781</idno><idno type="PII">S0167-4781(00)X0061-9</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999"></date><biblScope unit="volume">1445</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="86">86</biblScope><biblScope unit="page" to="98">98</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1999</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: The interaction of HIV-1 Tat protein with its recognition sequence, the trans-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-O-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-O-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>HIV-1</term></item><item><term>Tat protein</term></item><item><term>TAR RNA</term></item><item><term>Oligonucleotide</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1999">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><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:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity><istex:entity SYSTEM="gr3" NDATA="IMAGE" name="gr3"></istex:entity><istex:entity SYSTEM="gr4" NDATA="IMAGE" name="gr4"></istex:entity><istex:entity SYSTEM="gr5" NDATA="IMAGE" name="gr5"></istex:entity><istex:entity SYSTEM="gr6" NDATA="IMAGE" name="gr6"></istex:entity><istex:entity SYSTEM="gr7" NDATA="IMAGE" name="gr7"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>BBAEXP</jid><aid>93253</aid><ce:pii>S0167-4781(99)00019-6</ce:pii><ce:doi>10.1016/S0167-4781(99)00019-6</ce:doi><ce:copyright type="full-transfer" year="1999">Elsevier Science B.V.</ce:copyright></item-info><head><ce:title>Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the <ce:italic>trans</ce:italic>-activation responsive region (TAR) of HIV RNA</ce:title><ce:author-group><ce:author><ce:given-name>Béatrice</ce:given-name><ce:surname>Mestre</ce:surname><ce:cross-ref refid="FN1"><ce:sup>1</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Andrey</ce:given-name><ce:surname>Arzumanov</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Mohinder</ce:given-name><ce:surname>Singh</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Florence</ce:given-name><ce:surname>Boulmé</ce:surname><ce:cross-ref refid="FN2"><ce:sup>2</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Simon</ce:given-name><ce:surname>Litvak</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Michael J</ce:given-name><ce:surname>Gait</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="CORR1">*</ce:cross-ref><ce:e-address>mgait@mrc-lmb.cam.ac.uk</ce:e-address></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</ce:textfn></ce:affiliation><ce:correspondence id="CORR1"><ce:label>*</ce:label><ce:text>Corresponding author. Fax: +44 (1223) 402070</ce:text></ce:correspondence><ce:footnote id="FN1"><ce:label>1</ce:label><ce:note-para>Present address: Laboratoire de Biophysique Moléculaire, INSERM U-386, Université Victor Ségalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux, France.</ce:note-para></ce:footnote><ce:footnote id="FN2"><ce:label>2</ce:label><ce:note-para>Present address: Institute for Molecular Biology, Dr. Bohrgasse 9, 1030 Vienna, Austria.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="27" month="11" year="1998"></ce:date-received><ce:date-accepted day="27" month="1" year="1999"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>The interaction of HIV-1 Tat protein with its recognition sequence, the <ce:italic>trans</ce:italic>-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-<ce:italic>O</ce:italic>-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-<ce:italic>O</ce:italic>-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>HIV-1</ce:text></ce:keyword><ce:keyword><ce:text>Tat protein</ce:text></ce:keyword><ce:keyword><ce:text>TAR RNA</ce:text></ce:keyword><ce:keyword><ce:text>Oligonucleotide</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the trans -activation responsive region (TAR) of HIV RNA</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Oligonucleotide inhibition of the interaction of HIV-1 Tat protein with the</title></titleInfo><name type="personal"><namePart type="given">Béatrice</namePart><namePart type="family">Mestre</namePart><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation><description>Present address: Laboratoire de Biophysique Moléculaire, INSERM U-386, Université Victor Ségalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux, France.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Andrey</namePart><namePart type="family">Arzumanov</namePart><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Mohinder</namePart><namePart type="family">Singh</namePart><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Florence</namePart><namePart type="family">Boulmé</namePart><affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</affiliation><description>Present address: Institute for Molecular Biology, Dr. Bohrgasse 9, 1030 Vienna, Austria.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Simon</namePart><namePart type="family">Litvak</namePart><affiliation>EP-630, CNRS-Université Victor Ségalen Bordeaux 2, 1 rue Camille Saint Saëns, 33077 Bordeaux Cedex, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Michael J</namePart><namePart type="family">Gait</namePart><affiliation>E-mail: mgait@mrc-lmb.cam.ac.uk</affiliation><affiliation>Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK</affiliation><description>Corresponding author. Fax: +44 (1223) 402070</description><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">1999</dateIssued><copyrightDate encoding="w3cdtf">1999</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: The interaction of HIV-1 Tat protein with its recognition sequence, the trans-activation responsive region TAR is a potential target for drug discovery against HIV infection. We show by use of an in vitro competition filter binding interference assay that synthetic oligodeoxyribonucleotides complementary to the HIV-1 TAR RNA apical stem-loop and bulge region inhibit the binding of Tat protein or a Tat peptide (residues 37–72) better than two small molecules that have been shown to bind TAR RNA, Hoechst 33258 and neomycin B. The inhibition is not sensitive to length between 13 and 16 residues or precise positioning but shorter oligonucleotides are less effective. Enhanced inhibition was obtained for a 16-mer 2′-O-methyl oligoribonucleotide but not for C5-propyne pyrimidine-substituted oligonucleotides. Control non-antisense oligonucleotides were occasionally also effective in filter binding interference but only the complementary antisense 2′-O-methyl oligoribonucleotide was effective in gel mobility shift assays in direct TAR binding or in interference with Tat peptide binding to the TAR stem-loop. This is the first demonstration of effective inhibition of the Tat-TAR interaction by nuclease-stabilized oligonucleotide analogues.</abstract><note type="content">Fig. 1: (a) Regions of the HIV-1 Tat (BRU) protein and sequence of the Tat peptide (37–72). (b,c) Structures of the apical region of the HIV-1 TAR (BRU) and (MAL) stem-loops respectively as prepared synthetically, TAR BRU 39 and TAR MAL 38. The strain differences in the sequences of the loop and bulge region are boxed. For ease of comparison, the numbering scheme is harmonized between strains by considering nucleotide 25 to have been deleted in MAL (Δ25).</note><note type="content">Fig. 2: Competition filter binding interference assays showing inhibition of filter retention of TAR BRU 39 by Tat peptide (37–72) as a function of inhibitor concentration: (a) Hoechst 33258, neomycin B and a 16-mer oligonucleotide complementary to residues 22–37 (16 BRU (22–37)), (b) oligonucleotides 16 BRU (21–36), 16 BRU (22–37) and 16 BRU (23–38), (c) oligonucleotides 16 BRU (21–36), 15 BRU (22–36), 14 BRU (23–36) and 13 BRU (24–36), and (d) oligonucleotides 13 BRU (24–36), 11 BRU (26–36) and 6 BRU (31–36).</note><note type="content">Fig. 3: Competition filter binding interference assays showing inhibition of filter retention as a function of inhibitor concentration (a) for TAR BRU 39/Tat peptide (37–72) by oligonucleotides 15 BRU (21–35), 15 BRU mismatched and 15 BRU scrambled and (b) for TAR MAL 38/Tat protein by oligonucleotides 16 MAL (21–37), 16 MAL mismatched and 16 MAL scrambled.</note><note type="content">Fig. 4: Filter retention of 32P-labelled 16 BRU (23–38) oligonucleotide (a) as a function of concentration in the presence or absence of TAR BRU 39 and/or Tat peptide (37–72), and (b) as a function of increasing Tat peptide (37–72) concentration.</note><note type="content">Fig. 5: Competition filter binding interference assays showing inhibition of filter retention. (a) TAR BRU 39/Tat peptide (37–72) by oligonucleotides containing C5-propyne pyrimidines complementary to the loop (15 BRU L4), complementary to the stem (15 BRU S3), or both (15 BRU L4/S3) and unmodified oligonucleotide 15 BRU (21–35). The positions of C5-propyne incorporation are shown below underlined. (b) TAR MAL 38/Tat peptide (37–72) by all 2′-O-methyl oligoribonucleotides 16 MAL (21–37) OMe, 16 MAL mismatched OMe and 16 MAL scrambled (OMe) as well as unmodified 16 MAL (21–37) oligodeoxyribonucleotide.</note><note type="content">Fig. 6: Polyacrylamide gel mobility shift assay of binding of 2′-O-methyl oligoribonucleotides to a 274-residue 32P-labelled HIV-1 MAL transcript containing the full TAR RNA stem-loop. Lanes: 1, transcript alone; 2–6, 16 MAL OMe scrambled; 7–11, 16 MAL OMe mismatched; 12–16, 16 MAL OMe (21–37). The sequences of the oligonucleotides are shown in the lower part of Fig. 5.</note><note type="content">Fig. 7: Polyacrylamide gel mobility shift competition assay. Lanes: 1, 32P-labelled TAR MAL 38 alone; 2, TAR MAL 38+Tat peptide (37–72); 3, TAR MAL 38+16 MAL OMe (21–37); 4–6, TAR MAL 38, Tat peptide and increasing concentrations of 16 MAL OMe; 7, TAR MAL 38+mismatched OMe oligonucleotide; 8–10, TAR MAL 38, Tat peptide+increasing concentrations of mismatched OMe oligonucleotide; 11, TAR MAL 38+ scrambled OMe oligonucleotide; 12–14, TAR MAL 38, Tat peptide+increasing concentrations of scrambled OMe oligonucleotide.</note><subject><genre>Keywords</genre><topic>HIV-1</topic><topic>Tat protein</topic><topic>TAR RNA</topic><topic>Oligonucleotide</topic></subject><relatedItem type="host"><titleInfo><title>BBA - Gene Structure and Expression</title></titleInfo><titleInfo type="abbreviated"><title>BBAEXP</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1999</dateIssued></originInfo><identifier type="ISSN">0167-4781</identifier><identifier type="PII">S0167-4781(00)X0061-9</identifier><part><date>1999</date><detail type="volume"><number>1445</number><caption>vol.</caption></detail><detail type="issue"><number>1</number><caption>no.</caption></detail><extent unit="issue-pages"><start>1</start><end>183</end></extent><extent unit="pages"><start>86</start><end>98</end></extent></part></relatedItem><identifier type="istex">282A80C882C9652F58B797865503362587A61995</identifier><identifier type="ark">ark:/67375/6H6-7261QRW9-Z</identifier><identifier type="DOI">10.1016/S0167-4781(99)00019-6</identifier><identifier type="PII">S0167-4781(99)00019-6</identifier><accessCondition type="use and reproduction" contentType="copyright">©1999 Elsevier Science B.V.</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource><recordOrigin>Elsevier Science B.V., ©1999</recordOrigin></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-7261QRW9-Z/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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