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Nucleic Acids Research, 1988, Vol. 16, No. 24 11737-11757
© 1988

CHEMISTRY

Energy-structure correlations of plasmid DNA in different topological forms

Werner Thumm, Angela Seidl and Hans-Jürgen Hinz*

Institut für Biophysik und Physikalische Biochemie der Universität Regensburg, Universitätstraße 31, D-8400 Regensburg, FRG

*To whom correspondence should be addressed

Received September 29, 1988. Revised November 21, 1988. Accepted November 21, 1988.

Differential scanning microcalorimetry (DSC), UV absorption and circular dichroism (CD) have been used to study structure and stability of linear (lin), open circular (oc), supercoiled (cd) and relaxed circular duplex (rd) DNA and calf thymus (CT) DNA. Investigations were made in low salt buffer and in the presence of 7.2 M NaClO4 The chaotropic action of perchlorate promotes a reduction of the overall stability of DNA, which permits a direct determination of the transition enthalpies of all four DNA configurations. The stabilities against thermal denaturation have been found to increase in the series lin~oc<cd<rd. These relative stabilities can be rationalized on the basis of the linkage between supercoiling and secondary structural changes in topologically constrained duplex DNA. On the basis of these studies, a model of the melting process could be suggested that is consistent with the energetic and spectroscopic data.


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