Fine mapping of inherent flexibility variation along DNA molecules: validation by atomic force microscopy (AFM) in buffer.
Curvature and flexibility are structural properties of central importance to genome function. However, due to the difficulties in finding suitable experimental conditions, methods for studying one without the interference of the other have proven to be difficult. We propose a new approach that provides a measure of inherent flexibility of DNA by taking advantage of two powerful techniques, X-ray crystallography and nuclear magnetic resonance. Both techniques are able to detect local curvature on DNA fragments but, while the first analyzes DNA in the solid state, the second works on DNA in solution. Comparison of the two data sets allowed us to calculate the relative contribution to flexibility of the three rotations and three translations, which relate successive base pair planes for the ten different dinucleotide steps. These values were then used to compute the variation of flexibility along a given nucleotide sequence. This allowed us to validate the method experimentally through comparisons with maps of local fluctuations in DNA molecule trajectory constructed from atomic force microscopy imaging in solution. We conclude that the six dinucleotide-step parameters defined here provide a powerful tool for the exploration of DNA structure and, consequently will make an important contribution to our understanding of DNA-sequence-dependent biological processes.
Marilley
Monique
M
Laboratoire de régulation génique et fonctionnelle & microscopie champ proche (RGFCP), EA 3290, IFR 125, Faculté de Médecine, Réseau AFM Biologie, Université de la Méditerranée, 13385, Marseille cedex 5, France. Monique.Marilley@medecine.univ-mrs.fr
Sanchez-Sevilla
Albert
A
Rocca-Serra
José
J
D001483:Base Sequence; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry*; D003851:Deoxyribonucleases / Q000737:chemistry; D019281:Dimerization; D018625:Microscopy, Atomic Force / Q000379:methods*; D009038:Motion; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry; D009711:Nucleotides / Q000737:chemistry* / Q000378:metabolism; D018583:Pliability; D015203:Reproducibility of Results
D009707:Nucleosomes; D009711:Nucleotides; D004247:DNA; D003851:Deoxyribonucleases