Length-dependent energetics of (CTG)n and (CAG)n trinucleotide repeats
| Autor: | Jean-Louis Mergny, Samir Amrane, Horst H. Klump, Madhu Chauhan, Martin Mills, Barbara Saccà |
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| Rok vydání: | 2005 |
| Předmět: |
Circular dichroism
Enthalpy Calorimetry Biology Nucleic Acid Denaturation Oligomer Article chemistry.chemical_compound Differential scanning calorimetry Trinucleotide Repeats Genetics Base Sequence Circular Dichroism Temperature Endonucleases Footprinting Crystallography Oligodeoxyribonucleotides chemistry Biochemistry Intramolecular force Nucleic Acid Conformation Spectrophotometry Ultraviolet Biologie |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gki716 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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