Conformational fluctuations in single DNA molecules.
نویسندگان
چکیده
Measurement of fluorescent lifetimes of dye-tagged DNA molecules reveal the existence of different conformations. Conformational fluctuations observed by fluorescence correlation spectroscopy give rise to a relaxation behavior that is described by "stretched" exponentials and indicates the presence of a distribution of transition rates between two conformations. Whether this is an inhomogeneous distribution, where each molecule contributes with its own reaction rate to the overall distribution, or a homogeneous distribution, where the reaction rate of each molecule is time-dependent, is not yet known. We used a tetramethylrhodamine-linked 217-bp DNA oligonucleotide as a probe for conformational fluctuations. Fluorescence fluctuations from single DNA molecules attached to a streptavidin-coated surface directly show the transitions between two conformational states. The conformational fluctuations typical for single molecules are similar to those seen in single ion channels in cell membranes.
منابع مشابه
Single-molecule diffusion and conformational dynamics by spatial integration of temporal fluctuations
Single-molecule localization and tracking has been used to translate spatiotemporal information of individual molecules to map their diffusion behaviours. However, accurate analysis of diffusion behaviours and including other parameters, such as the conformation and size of molecules, remain as limitations to the method. Here, we report a method that addresses the limitations of existing single...
متن کاملColloidal Particle Motion as a Diagnostic of DNA Conformational Transitions
Tethered particle motion is an experimental technique to monitor conformational changes in single molecules of DNA in real time, by observing the position fluctuations of a micrometersize particle attached to the DNA. This article reviews some recent work on theoretical problems inherent in the interpretation of TPM experiments, both in equilibrium and dynamical aspects.
متن کاملCharacterizing single-molecule FRET dynamics with probability distribution analysis.
Probability distribution analysis (PDA) is a recently developed statistical tool for predicting the shapes of single-molecule fluorescence resonance energy transfer (smFRET) histograms, which allows the identification of single or multiple static molecular species within a single histogram. We used a generalized PDA method to predict the shapes of FRET histograms for molecules interconverting d...
متن کاملPrincipal-components analysis of shape fluctuations of single DNA molecules.
Thermal fluctuations agitate molecules in solution over a broad range of times and distances. By passively watching the shape fluctuations of a thermally driven biomolecule, one can infer properties of the underlying interactions that determine the motion. We applied this concept to single molecules of fluorescently labeled lambda-DNA, a key model system for polymer physics. In contrast to most...
متن کاملConformational fluctuations of DNA helix.
Treatment of a homogeneous sample of circular DNA molecules that contain at least one single-strand scission per molecule with ligase yields a population of covalently closed circles heterogeneous in the topological winding number alpha. Under proper conditions, the product molecules with different values of alpha can all be resolved by gel electrophoresis. The distributions in alpha for severa...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Proceedings of the National Academy of Sciences of the United States of America
دوره 94 20 شماره
صفحات -
تاریخ انتشار 1997