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Nucleic Acids Research, 1984, Vol. 12, No. 7 3245-3256
© 1984

CHEMISTRY

Deoxydodecanucleotide heteroduplex d(TTTTATAATAAA). d(TTTATTATAAAA) containing the promoter Pribnow sequence TATAAT. I. Double-helix stability by UV spectrophotometry and calorimetry

Krystyna Bolewska, Anna Zielenkiewicz+ and Kazimierz L. Wierzchowski*

Institute of Biochemistry and Biophysics, Polish Academy of Sciences 02-532 Warszawa +Institute of Physical Chemistry, Polish Academy of Sciences 01-224 Warszawa, Poland

*To whom all correspondence should be addressed: c/o Prof.B.Pullman, Institut de Biologie Physico-Chimique, 13, rue Pierre et Marie Curie, 75005 Paris, France

Received February 7, 1984. Accepted March 13, 1984.

Thermally induced structural transition in the d(TTTTATAATAAA) d(TTTATTATAAAA) heteroduplex is characterized by UV–spectroscopy and differential scanning calorimetry. At low salt(<0.1 M)the occurrence of a cooperative transition in the lower temperature range, followed by a broad transition connected with small increase in absorbance is observed. At high salt (≥0.2 M)a single, monophasic transition appears. Linear dependence of the latter on log of salt concentration (dTm:dlogM=14.2°C) and of l/Tm on log of oligomer concentration [derived therefrom {delta}H (v.H.) =77.1 kcal/mole (duplex)]allows relating it to the melting of the heteroduplex helix. The non-cooperative transition, independent of oligomer concentration and similar to that of the single chain, was attributed to melting of short hairpin helices upon heteroduplex dissociation. Calorimetric enthalpy: 75.6 kcal/ /mole(duplex)proved significantly lower than predicted from known calorimetric data for poly [d(AT)] and poly d(A)·poly d(T).


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