A study of the interactions that stabilize DNA frayed wires
| Autor: | Robert B. Macgregor, Amr S. Helmy, Rashid Abu-Ghazalah, J. Irizar |
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| Rok vydání: | 2009 |
| Předmět: |
Gel electrophoresis
Circular dichroism Guanine analog Guanine Stereochemistry Organic Chemistry Biophysics Temperature DNA Methylation G-quadruplex Nucleic Acid Denaturation Biochemistry chemistry.chemical_compound Crystallography chemistry Oligodeoxyribonucleotides Deoxyguanosine Nucleic Acid Conformation Thermodynamics A-DNA DNA |
| Zdroj: | Biophysical chemistry. 147(3) |
| ISSN: | 1873-4200 |
| Popis: | Oligodeoxyribonucleotides (ODNs) with long, terminal runs of consecutive guanines, and either a dA or dT tract at the other end form higher-order structures called DNA frayed wires. These aggregates self-assemble into species consisting of 2, 3, 4, 5, … associated strands. Some of the remarkable features of these structures are their extreme thermostability and resistance to chemical denaturants and nucleases. However, the nature of the molecular interactions that stabilize these structures remains unclear. Based on dimethyl sulfate (DMS) methylation results, our group previously proposed DNA frayed wires to be a unique set of nucleic-acid assemblies in which the N7 of guanine does not participate in the guanine–guanine interactions. To probe the hydrogen bonding involved in the stabilization of d(A 15 G 15 ) frayed wires, we used Raman spectroscopy in which the DNA sample is held in photonic crystal fibers. This technique significantly enhances the signals thus allowing the use of very low laser power. Based on our results for d(A 15 G 15 ) and those of incorporating the isoelectronic guanine analog pyrazolo[3,4,-d]pyrimidine or PPG, into a frayed wire-forming sequence, we provide evidence that these structures are based on the G-quadruplex model. Furthermore, from the Raman spectrum, we observed markers that are consistent with the presence of deoxyguanosine residues in the syn conformation, this suggests the presence of anti-parallel G-quadruplexes. To identify the species that contain syn guanine residues, we used circular dichroism and gel electrophoresis to study an ODN in which all of the guanine residues were brominated, d(A 15 8-Br G 15 ). In the presence of potassium, d(A 15 8-Br G 15 ) forms what appears to be an anti-parallel dimeric G-quadruplex. To our knowledge, this is the first report of a DNA sequence having all its guanine residues replaced by 8-bromo-guanine and maintaining its ability to form a G-quadruplex structure. |
| Databáze: | OpenAIRE |
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