Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.
Identifieur interne : 002840 ( Ncbi/Merge ); précédent : 002839; suivant : 002841Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.
Auteurs : D. Praseuth [France] ; L. Perrouault ; T. Le Doan ; M. Chassignol ; N. Thuong ; C. HélèneSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1988.
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Abstract
A photocrosslinking reagent (p-azidophenacyl) was covalently linked to an octathymidylate synthesized with either the natural (beta) anomer of thymidine or the synthetic (alpha) anomer. The oligothymidylate was further substituted by an acridine derivative to stabilize the hybrid formed with a complementary octadeoxyadenylate sequence via intercalation. A single-stranded 27-mer containing a (dA)8 sequence and a 27-mer duplex containing a (dA.dT)8 sequence were used as targets. Upon UV irradiation, photocrosslinking of the octathymidylate to its target sequence was observed, generating bands that migrated more slowly in denaturing gels. In the 27-mer duplex, both strands were photocrosslinked to the octathymidylate. Upon alkaline treatment of the irradiated samples, cleavage of the 27-mers was observed at specific sites. These reactions were analyzed at different salt concentrations. The location of the cleavage sites allowed us to demonstrate the following. (i) Both alpha and beta oligothymidylates can recognize a DNA double helix containing an oligo(dA).oligo(dT) sequence; the oligothymidylate binds to the major groove of DNA in a parallel orientation with respect to the adenine-containing strand of the DNA double helix. (ii) alpha oligothymidylates form helices with a complementary single-stranded oligodeoxyadenylate; the two strands have a parallel orientation independently of whether or not an intercalating agent is attached to the oligothymidylate. (iii) At low salt concentration, beta oligothymidylates form a double helix with an oligodeoxyadenylate in which, as expected, the two strands are antiparallel; at high salt concentration, a triple helix is formed in which the second oligothymidylate is oriented parallel to the adenine-containing strand. These results show that it is possible to recognize an oligopurine.oligopyrimidine sequence in a DNA double helix via local triple-helix formation and to target photochemical reactions to specific sequences in both double-stranded and single-stranded nucleic acids.
DOI: 10.1073/pnas.85.5.1349
PubMed: 3422738
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.</title><author><name sortKey="Praseuth, D" sort="Praseuth, D" uniqKey="Praseuth D" first="D" last="Praseuth">D. Praseuth</name><affiliation wicri:level="3"><nlm:affiliation>Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Perrouault, L" sort="Perrouault, L" uniqKey="Perrouault L" first="L" last="Perrouault">L. Perrouault</name></author><author><name sortKey="Le Doan, T" sort="Le Doan, T" uniqKey="Le Doan T" first="T" last="Le Doan">T. Le Doan</name></author><author><name sortKey="Chassignol, M" sort="Chassignol, M" uniqKey="Chassignol M" first="M" last="Chassignol">M. Chassignol</name></author><author><name sortKey="Thuong, N" sort="Thuong, N" uniqKey="Thuong N" first="N" last="Thuong">N. Thuong</name></author><author><name sortKey="Helene, C" sort="Helene, C" uniqKey="Helene C" first="C" last="Hélène">C. 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Praseuth</name><affiliation wicri:level="3"><nlm:affiliation>Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Perrouault, L" sort="Perrouault, L" uniqKey="Perrouault L" first="L" last="Perrouault">L. Perrouault</name></author><author><name sortKey="Le Doan, T" sort="Le Doan, T" uniqKey="Le Doan T" first="T" last="Le Doan">T. Le Doan</name></author><author><name sortKey="Chassignol, M" sort="Chassignol, M" uniqKey="Chassignol M" first="M" last="Chassignol">M. Chassignol</name></author><author><name sortKey="Thuong, N" sort="Thuong, N" uniqKey="Thuong N" first="N" last="Thuong">N. Thuong</name></author><author><name sortKey="Helene, C" sort="Helene, C" uniqKey="Helene C" first="C" last="Hélène">C. Hélène</name></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><imprint><date when="1988" type="published">1988</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cross-Linking Reagents</term><term>DNA</term><term>Nucleic Acid Conformation</term><term>Oligodeoxyribonucleotides</term><term>Photochemistry</term><term>Structure-Activity Relationship</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN</term><term>Conformation d'acide nucléique</term><term>Oligodésoxyribonucléotides</term><term>Photochimie</term><term>Relation structure-activité</term><term>Réactifs réticulants</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Cross-Linking Reagents</term><term>DNA</term><term>Oligodeoxyribonucleotides</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Nucleic Acid Conformation</term><term>Photochemistry</term><term>Structure-Activity Relationship</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN</term><term>Conformation d'acide nucléique</term><term>Oligodésoxyribonucléotides</term><term>Photochimie</term><term>Relation structure-activité</term><term>Réactifs réticulants</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A photocrosslinking reagent (p-azidophenacyl) was covalently linked to an octathymidylate synthesized with either the natural (beta) anomer of thymidine or the synthetic (alpha) anomer. The oligothymidylate was further substituted by an acridine derivative to stabilize the hybrid formed with a complementary octadeoxyadenylate sequence via intercalation. A single-stranded 27-mer containing a (dA)8 sequence and a 27-mer duplex containing a (dA.dT)8 sequence were used as targets. Upon UV irradiation, photocrosslinking of the octathymidylate to its target sequence was observed, generating bands that migrated more slowly in denaturing gels. In the 27-mer duplex, both strands were photocrosslinked to the octathymidylate. Upon alkaline treatment of the irradiated samples, cleavage of the 27-mers was observed at specific sites. These reactions were analyzed at different salt concentrations. The location of the cleavage sites allowed us to demonstrate the following. (i) Both alpha and beta oligothymidylates can recognize a DNA double helix containing an oligo(dA).oligo(dT) sequence; the oligothymidylate binds to the major groove of DNA in a parallel orientation with respect to the adenine-containing strand of the DNA double helix. (ii) alpha oligothymidylates form helices with a complementary single-stranded oligodeoxyadenylate; the two strands have a parallel orientation independently of whether or not an intercalating agent is attached to the oligothymidylate. (iii) At low salt concentration, beta oligothymidylates form a double helix with an oligodeoxyadenylate in which, as expected, the two strands are antiparallel; at high salt concentration, a triple helix is formed in which the second oligothymidylate is oriented parallel to the adenine-containing strand. These results show that it is possible to recognize an oligopurine.oligopyrimidine sequence in a DNA double helix via local triple-helix formation and to target photochemical reactions to specific sequences in both double-stranded and single-stranded nucleic acids.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">3422738</PMID><DateCompleted><Year>1988</Year><Month>04</Month><Day>01</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>01</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0027-8424</ISSN><JournalIssue CitedMedium="Print"><Volume>85</Volume><Issue>5</Issue><PubDate><Year>1988</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Proceedings of the National Academy of Sciences of the United States of America</Title><ISOAbbreviation>Proc. Natl. Acad. Sci. U.S.A.</ISOAbbreviation></Journal><ArticleTitle>Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.</ArticleTitle><Pagination><MedlinePgn>1349-53</MedlinePgn></Pagination><Abstract><AbstractText>A photocrosslinking reagent (p-azidophenacyl) was covalently linked to an octathymidylate synthesized with either the natural (beta) anomer of thymidine or the synthetic (alpha) anomer. The oligothymidylate was further substituted by an acridine derivative to stabilize the hybrid formed with a complementary octadeoxyadenylate sequence via intercalation. A single-stranded 27-mer containing a (dA)8 sequence and a 27-mer duplex containing a (dA.dT)8 sequence were used as targets. Upon UV irradiation, photocrosslinking of the octathymidylate to its target sequence was observed, generating bands that migrated more slowly in denaturing gels. In the 27-mer duplex, both strands were photocrosslinked to the octathymidylate. Upon alkaline treatment of the irradiated samples, cleavage of the 27-mers was observed at specific sites. These reactions were analyzed at different salt concentrations. The location of the cleavage sites allowed us to demonstrate the following. (i) Both alpha and beta oligothymidylates can recognize a DNA double helix containing an oligo(dA).oligo(dT) sequence; the oligothymidylate binds to the major groove of DNA in a parallel orientation with respect to the adenine-containing strand of the DNA double helix. (ii) alpha oligothymidylates form helices with a complementary single-stranded oligodeoxyadenylate; the two strands have a parallel orientation independently of whether or not an intercalating agent is attached to the oligothymidylate. (iii) At low salt concentration, beta oligothymidylates form a double helix with an oligodeoxyadenylate in which, as expected, the two strands are antiparallel; at high salt concentration, a triple helix is formed in which the second oligothymidylate is oriented parallel to the adenine-containing strand. These results show that it is possible to recognize an oligopurine.oligopyrimidine sequence in a DNA double helix via local triple-helix formation and to target photochemical reactions to specific sequences in both double-stranded and single-stranded nucleic acids.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Praseuth</LastName><ForeName>D</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, Paris, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Perrouault</LastName><ForeName>L</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Le Doan</LastName><ForeName>T</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Chassignol</LastName><ForeName>M</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Thuong</LastName><ForeName>N</ForeName><Initials>N</Initials></Author><Author ValidYN="Y"><LastName>Hélène</LastName><ForeName>C</ForeName><Initials>C</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Proc Natl Acad Sci U S A</MedlineTA><NlmUniqueID>7505876</NlmUniqueID><ISSNLinking>0027-8424</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D003432">Cross-Linking Reagents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009838">Oligodeoxyribonucleotides</NameOfSubstance></Chemical><Chemical><RegistryNumber>9007-49-2</RegistryNumber><NameOfSubstance UI="D004247">DNA</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName 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last="Chassignol">M. Chassignol</name><name sortKey="Helene, C" sort="Helene, C" uniqKey="Helene C" first="C" last="Hélène">C. Hélène</name><name sortKey="Le Doan, T" sort="Le Doan, T" uniqKey="Le Doan T" first="T" last="Le Doan">T. Le Doan</name><name sortKey="Perrouault, L" sort="Perrouault, L" uniqKey="Perrouault L" first="L" last="Perrouault">L. Perrouault</name><name sortKey="Thuong, N" sort="Thuong, N" uniqKey="Thuong N" first="N" last="Thuong">N. Thuong</name></noCountry><country name="France"><region name="Île-de-France"><name sortKey="Praseuth, D" sort="Praseuth, D" uniqKey="Praseuth D" first="D" last="Praseuth">D. Praseuth</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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