Length-dependent energetics of (CTG)n and (CAG)n trinucleotide repeats.
Trinucleotide repeats are involved in a number of debilitating diseases such as myotonic dystrophy. Twelve to seventy-five base-long (CTG)n oligodeoxynucleotides were analysed using a combination of biophysical [UV-absorbance, circular dichroism and differential scanning calorimetry (DSC)] and biochemical methods (non-denaturing gel electrophoresis and enzymatic footprinting). All oligomers formed stable intramolecular structures under near physiological conditions with a melting temperature that was only weakly dependent on oligomer length. Thermodynamic analysis of the denaturation process by UV-melting and calorimetric experiments revealed an unprecedented length-dependent discrepancy between the enthalpy values deduced from model-dependent (UV-melting) and model-independent (calorimetry) experiments. Evidence for non-zero molar heat capacity changes was also derived from the analysis of the Arrhenius plots and DSC profiles. Such behaviour is analysed in the framework of an intramolecular 'branched-hairpin' model, in which long CTG oligomers do not fold into a simple long hairpin-stem intramolecular structure, but allow the formation of several independent folding units of unequal stability. We demonstrate that, for sequences ranging from 12 to 25 CTG repeats, an intramolecular structure with two loops is formed which we will call 'bis-hairpin'. Similar results were also found for CAG oligomers, suggesting that this observation may be extended to various trinucleotide repeats-containing sequences.
Amrane
Samir
S
Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, USM 503 INSERM UR 565, CNRS UMR 8646, 43 rue Cuvier, 75231 Paris cedex 05, France.
Saccà
Barbara
B
Mills
Martin
M
Chauhan
Madhu
M
Klump
Horst H
HH
Mergny
Jean-Louis
JL
D001483:Base Sequence; D002151:Calorimetry; D002942:Circular Dichroism; D004720:Endonucleases / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D018911:Trinucleotide Repeats
D009838:Oligodeoxyribonucleotides; D004720:Endonucleases