MersV1, Ncbi, Merge, bibRecord, 000666

Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures.

Identifieur interne : 000666 ( Ncbi/Merge ); précédent : 000665; suivant : 000667

Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures.

Auteurs : Jun-Huei Fan [États-Unis] ; Elena Bochkareva ; Alexey Bochkarev ; Donald M. Gray

Source :

Biochemistry [ 1520-4995 ] ; 2009.

RBID : pubmed:19187036

Descripteurs français

English descriptors

KwdEn :
- Circular Dichroism, Crystallography, X-Ray, Electrophoretic Mobility Shift Assay, G-Quadruplexes, Hot Temperature, Humans, Nucleic Acid Conformation, Protein Binding (genetics), Protein Stability, Replication Protein A (chemistry), Replication Protein A (genetics), Replication Protein A (metabolism).
MESH :
- chemical , chemistry : Replication Protein A.
- genetics : Protein Binding, Replication Protein A.
- chemical , metabolism : Replication Protein A.
- Circular Dichroism, Crystallography, X-Ray, Electrophoretic Mobility Shift Assay, G-Quadruplexes, Hot Temperature, Humans, Nucleic Acid Conformation, Protein Stability.

Abstract

Noncanonical DNA structures such as G-quadruplexes might obstruct the binding of hRPA, compromising the accuracy of replication, and be a source of genomic instability. In this study, circular dichroism (CD) and electrophoretic mobility shift assay (EMSA) experiments were used to show that hRPA can bind and melt nontelomeric, intramolecular DNA G-quadruplexes under physiologically germane conditions. EMSA results show that hRPA binds to a 58-mer that includes an embedded quadruplex with an affinity equal to or greater than to nonquadruplex forming 58-mers. Moreover, hRPA binds to a 26-mer purine-rich quadruplex-forming sequence with an affinity indistinguishable from that for binding to the complementary pyrimidine-rich sequence. Under the same conditions, hRPA does not have significant affinity for binding to the duplex formed from the two sequences. Thus, DNA secondary structures can significantly modulate the binding affinity of hRPA over and above its known preference for pyrimidine-rich single-stranded sequences, so that at least some intramolecular G-quadruplex structures may not inhibit hRPA binding during DNA replication. CD spectral changes in combination with EMSA titrations suggest that one hRPA heterotrimer is sufficient to form a stable complex with an unfolded 26-mer G-quadruplex prior to the binding of a second hRPA molecule.

DOI: 10.1021/bi801538h
PubMed: 19187036

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Le document en format XML

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<author><name sortKey="Fan, Jun Huei" sort="Fan, Jun Huei" uniqKey="Fan J" first="Jun-Huei" last="Fan">Jun-Huei Fan</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Molecular and Cell Biology, Mail Stop FO31, The University of Texas at Dallas, 800 West Campbell Road, Richardson, Texas 75080, USA.</nlm:affiliation>
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<wicri:regionArea>Department of Molecular and Cell Biology, Mail Stop FO31, The University of Texas at Dallas, 800 West Campbell Road, Richardson, Texas 75080</wicri:regionArea>
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<term>G-Quadruplexes</term>
<term>Hot Temperature</term>
<term>Humans</term>
<term>Nucleic Acid Conformation</term>
<term>Protein Binding (genetics)</term>
<term>Protein Stability</term>
<term>Replication Protein A (chemistry)</term>
<term>Replication Protein A (genetics)</term>
<term>Replication Protein A (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Conformation d'acide nucléique</term>
<term>Cristallographie aux rayons X</term>
<term>Dichroïsme circulaire</term>
<term>G-quadruplexes</term>
<term>Humains</term>
<term>Liaison aux protéines (génétique)</term>
<term>Protéine A de réplication ()</term>
<term>Protéine A de réplication (génétique)</term>
<term>Protéine A de réplication (métabolisme)</term>
<term>Stabilité protéique</term>
<term>Température élevée</term>
<term>Test de retard de migration électrophorétique</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Replication Protein A</term>
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<term>Replication Protein A</term>
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<term>Protéine A de réplication</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Replication Protein A</term>
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</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Circular Dichroism</term>
<term>Crystallography, X-Ray</term>
<term>Electrophoretic Mobility Shift Assay</term>
<term>G-Quadruplexes</term>
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<term>Humans</term>
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<term>Protein Stability</term>
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<term>Cristallographie aux rayons X</term>
<term>Dichroïsme circulaire</term>
<term>G-quadruplexes</term>
<term>Humains</term>
<term>Protéine A de réplication</term>
<term>Stabilité protéique</term>
<term>Température élevée</term>
<term>Test de retard de migration électrophorétique</term>
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<front><div type="abstract" xml:lang="en">Noncanonical DNA structures such as G-quadruplexes might obstruct the binding of hRPA, compromising the accuracy of replication, and be a source of genomic instability. In this study, circular dichroism (CD) and electrophoretic mobility shift assay (EMSA) experiments were used to show that hRPA can bind and melt nontelomeric, intramolecular DNA G-quadruplexes under physiologically germane conditions. EMSA results show that hRPA binds to a 58-mer that includes an embedded quadruplex with an affinity equal to or greater than to nonquadruplex forming 58-mers. Moreover, hRPA binds to a 26-mer purine-rich quadruplex-forming sequence with an affinity indistinguishable from that for binding to the complementary pyrimidine-rich sequence. Under the same conditions, hRPA does not have significant affinity for binding to the duplex formed from the two sequences. Thus, DNA secondary structures can significantly modulate the binding affinity of hRPA over and above its known preference for pyrimidine-rich single-stranded sequences, so that at least some intramolecular G-quadruplex structures may not inhibit hRPA binding during DNA replication. CD spectral changes in combination with EMSA titrations suggest that one hRPA heterotrimer is sufficient to form a stable complex with an unfolded 26-mer G-quadruplex prior to the binding of a second hRPA molecule.</div>
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<Abstract><AbstractText>Noncanonical DNA structures such as G-quadruplexes might obstruct the binding of hRPA, compromising the accuracy of replication, and be a source of genomic instability. In this study, circular dichroism (CD) and electrophoretic mobility shift assay (EMSA) experiments were used to show that hRPA can bind and melt nontelomeric, intramolecular DNA G-quadruplexes under physiologically germane conditions. EMSA results show that hRPA binds to a 58-mer that includes an embedded quadruplex with an affinity equal to or greater than to nonquadruplex forming 58-mers. Moreover, hRPA binds to a 26-mer purine-rich quadruplex-forming sequence with an affinity indistinguishable from that for binding to the complementary pyrimidine-rich sequence. Under the same conditions, hRPA does not have significant affinity for binding to the duplex formed from the two sequences. Thus, DNA secondary structures can significantly modulate the binding affinity of hRPA over and above its known preference for pyrimidine-rich single-stranded sequences, so that at least some intramolecular G-quadruplex structures may not inhibit hRPA binding during DNA replication. CD spectral changes in combination with EMSA titrations suggest that one hRPA heterotrimer is sufficient to form a stable complex with an unfolded 26-mer G-quadruplex prior to the binding of a second hRPA molecule.</AbstractText>
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Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures.

Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki