6 results on '"Micheletti, C."'
Search Results
2. Transcriptional supercoiling boosts topoisomerase II-mediated knotting of intracellular DNA.
- Author
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Valdés A, Coronel L, Martínez-García B, Segura J, Dyson S, Díaz-Ingelmo O, Micheletti C, and Roca J
- Subjects
- Chromatin ultrastructure, DNA Topoisomerases, Type I metabolism, DNA, Fungal metabolism, Humans, Nucleosomes metabolism, Saccharomyces cerevisiae metabolism, DNA Topoisomerases, Type II physiology, DNA, Superhelical metabolism, Nucleic Acid Conformation, Saccharomyces cerevisiae Proteins physiology, Transcription, Genetic genetics
- Abstract
Recent studies have revealed that the DNA cross-inversion mechanism of topoisomerase II (topo II) not only removes DNA supercoils and DNA replication intertwines, but also produces small amounts of DNA knots within the clusters of nucleosomes that conform to eukaryotic chromatin. Here, we examine how transcriptional supercoiling of intracellular DNA affects the occurrence of these knots. We show that although (-) supercoiling does not change the basal DNA knotting probability, (+) supercoiling of DNA generated in front of the transcribing complexes increases DNA knot formation over 25-fold. The increase of topo II-mediated DNA knotting occurs both upon accumulation of (+) supercoiling in topoisomerase-deficient cells and during normal transcriptional supercoiling of DNA in TOP1 TOP2 cells. We also show that the high knotting probability (Pkn ≥ 0.5) of (+) supercoiled DNA reflects a 5-fold volume compaction of the nucleosomal fibers in vivo. Our findings indicate that topo II-mediated DNA knotting could be inherent to transcriptional supercoiling of DNA and other chromatin condensation processes and establish, therefore, a new crucial role of topoisomerase II in resetting the knotting-unknotting homeostasis of DNA during chromatin dynamics., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2019
- Full Text
- View/download PDF
3. Dynamics of supercoiled DNA with complex knots: large-scale rearrangements and persistent multi-strand interlocking.
- Author
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Coronel L, Suma A, and Micheletti C
- Subjects
- DNA genetics, DNA metabolism, DNA, Superhelical genetics, DNA, Superhelical metabolism, Kinetics, Plasmids genetics, Plasmids metabolism, Time Factors, DNA chemistry, DNA, Superhelical chemistry, Molecular Dynamics Simulation, Nucleic Acid Conformation, Plasmids chemistry
- Abstract
Knots and supercoiling are both introduced in bacterial plasmids by catalytic processes involving DNA strand passages. While the effects on plasmid organization has been extensively studied for knotting and supercoiling taken separately, much less is known about their concurrent action. Here, we use molecular dynamics simulations and oxDNA, an accurate mesoscopic DNA model, to study the kinetic and metric changes introduced by complex (five-crossing) knots and supercoiling in 2 kbp-long DNA rings. We find several unexpected results. First, the conformational ensemble is dominated by two distinct states, differing in branchedness and knot size. Secondly, fluctuations between these states are as fast as the metric relaxation of unknotted rings. In spite of this, certain boundaries of knotted and plectonemically-wound regions can persist over much longer timescales. These pinned regions involve multiple strands that are interlocked by the cooperative action of topological and supercoiling constraints. Their long-lived character may be relevant for the simplifying action of topoisomerases.
- Published
- 2018
- Full Text
- View/download PDF
4. Spatial confinement induces hairpins in nicked circular DNA.
- Author
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Japaridze A, Orlandini E, Smith KB, Gmür L, Valle F, Micheletti C, and Dietler G
- Subjects
- Base Sequence genetics, DNA ultrastructure, DNA Breaks, Single-Stranded, DNA, Circular genetics, DNA, Circular ultrastructure, DNA-Binding Proteins genetics, DNA-Binding Proteins ultrastructure, Microscopy, Atomic Force, Models, Molecular, DNA chemistry, DNA, Circular chemistry, DNA-Binding Proteins chemistry, Nucleic Acid Conformation
- Abstract
In living cells, DNA is highly confined in space with the help of condensing agents, DNA binding proteins and high levels of supercoiling. Due to challenges associated with experimentally studying DNA under confinement, little is known about the impact of spatial confinement on the local structure of the DNA. Here, we have used well characterized slits of different sizes to collect high resolution atomic force microscopy images of confined circular DNA with the aim of assessing the impact of the spatial confinement on global and local conformational properties of DNA. Our findings, supported by numerical simulations, indicate that confinement imposes a large mechanical stress on the DNA as evidenced by a pronounced anisotropy and tangent-tangent correlation function with respect to non-constrained DNA. For the strongest confinement we observed nanometer sized hairpins and interwound structures associated with the nicked sites in the DNA sequence. Based on these findings, we propose that spatial DNA confinement in vivo can promote the formation of localized defects at mechanically weak sites that could be co-opted for biological regulatory functions., (© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2017
- Full Text
- View/download PDF
5. Elastic network models for RNA: a comparative assessment with molecular dynamics and SHAPE experiments.
- Author
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Pinamonti G, Bottaro S, Micheletti C, and Bussi G
- Subjects
- Molecular Dynamics Simulation, Nucleic Acid Conformation, Osmolar Concentration, Models, Molecular, RNA chemistry
- Abstract
Elastic network models (ENMs) are valuable and efficient tools for characterizing the collective internal dynamics of proteins based on the knowledge of their native structures. The increasing evidence that the biological functionality of RNAs is often linked to their innate internal motions poses the question of whether ENM approaches can be successfully extended to this class of biomolecules. This issue is tackled here by considering various families of elastic networks of increasing complexity applied to a representative set of RNAs. The fluctuations predicted by the alternative ENMs are stringently validated by comparison against extensive molecular dynamics simulations and SHAPE experiments. We find that simulations and experimental data are systematically best reproduced by either an all-atom or a three-beads-per-nucleotide representation (sugar-base-phosphate), with the latter arguably providing the best balance of accuracy and computational complexity., (© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2015
- Full Text
- View/download PDF
6. McGenus: a Monte Carlo algorithm to predict RNA secondary structures with pseudoknots.
- Author
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Bon M, Micheletti C, and Orland H
- Subjects
- Nucleic Acid Conformation, Algorithms, Monte Carlo Method, RNA chemistry
- Abstract
We present McGenus, an algorithm to predict RNA secondary structures with pseudoknots. The method is based on a classification of RNA structures according to their topological genus. McGenus can treat sequences of up to 1000 bases and performs an advanced stochastic search of their minimum free energy structure allowing for non-trivial pseudoknot topologies. Specifically, McGenus uses a Monte Carlo algorithm with replica exchange for minimizing a general scoring function which includes not only free energy contributions for pair stacking, loop penalties, etc. but also a phenomenological penalty for the genus of the pairing graph. The good performance of the stochastic search strategy was successfully validated against TT2NE which uses the same free energy parametrization and performs exhaustive or partially exhaustive structure search, albeit for much shorter sequences (up to 200 bases). Next, the method was applied to other RNA sets, including an extensive tmRNA database, yielding results that are competitive with existing algorithms. Finally, it is shown that McGenus highlights possible limitations in the free energy scoring function. The algorithm is available as a web server at http://ipht.cea.fr/rna/mcgenus.php.
- Published
- 2013
- Full Text
- View/download PDF
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