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The different binding modes of Hoechst 33258 to DNA studied by electirc linear dichroism

Identifieur interne : 004B14 ( Main/Exploration ); précédent : 004B13; suivant : 004B15

The different binding modes of Hoechst 33258 to DNA studied by electirc linear dichroism

Auteurs : Christian Bailly [Belgique] ; Pierre Colson ; Jean-Pierre Hénichart [Belgique] ; Claude Houssier

Source :

RBID : ISTEX:5ADDD26708757CE73A17DD3CF374A3C971112FFE

Abstract

The binding mode of the bisbenzimidazoie derivative Hoechst 33258 to a series of DNAs and polynucleotides has been investigated by electric linear dichroism. Positive reduced dichroisms were measured for the poly(dA-dT).poly(dA-dT)- and poly(dA).poly(dT)-Hoechst complexes in agreement with a deep penetration of the drug into the minor groove. Similarly, the drug displays positive reduced dichroism in the presence of the DNAs from calf thymus, Clostridium perfringens and Collphage T4. Conversely, negative reduced dichroisms were obtained when Hoechst 33258 was bound to poly(dG-dC).poly(dG-dC), poly(dAdC). poly(dG-dT) and poly(dG). poly(dC) as well as with the GC-rich DNA from Micrococcus lysodelkticus indicating that in this case minor groove binding cannot occur. Substitution of guanosines for inosines induces a reversal of the reduced dichroism from negative to positive. Therefore, as anticipated it is the 2-amino group of guanines protruding in this groove which prevents Hoechst 33258 from getting access to the minor groove of GC sequences. The ELD data obtained with the GC-rich biopolymers are consistent with an intercalative binding. Competition experiments performed with the intercalating drug proflavine lend credence to the involvement of an intercalative binding rather than to an external or major groove binding of Hoechst 33258 at GC sequences.

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DOI: 10.1093/nar/21.16.3705


Affiliations:


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<div type="abstract">The binding mode of the bisbenzimidazoie derivative Hoechst 33258 to a series of DNAs and polynucleotides has been investigated by electric linear dichroism. Positive reduced dichroisms were measured for the poly(dA-dT).poly(dA-dT)- and poly(dA).poly(dT)-Hoechst complexes in agreement with a deep penetration of the drug into the minor groove. Similarly, the drug displays positive reduced dichroism in the presence of the DNAs from calf thymus, Clostridium perfringens and Collphage T4. Conversely, negative reduced dichroisms were obtained when Hoechst 33258 was bound to poly(dG-dC).poly(dG-dC), poly(dAdC). poly(dG-dT) and poly(dG). poly(dC) as well as with the GC-rich DNA from Micrococcus lysodelkticus indicating that in this case minor groove binding cannot occur. Substitution of guanosines for inosines induces a reversal of the reduced dichroism from negative to positive. Therefore, as anticipated it is the 2-amino group of guanines protruding in this groove which prevents Hoechst 33258 from getting access to the minor groove of GC sequences. The ELD data obtained with the GC-rich biopolymers are consistent with an intercalative binding. Competition experiments performed with the intercalating drug proflavine lend credence to the involvement of an intercalative binding rather than to an external or major groove binding of Hoechst 33258 at GC sequences.</div>
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