Santiago Cuesta-López

TRF2 promotes, remodels and protects telomeric Holliday junctions

The ability of the telomeric DNA‐binding protein, TRF2, to stimulate t‐loop formation while preventing t‐loop deletion is believed to be crucial to maintain telomere integrity in mammals. However, little is known on the molecular mechanisms behind these properties of TRF2. In this report, we show that TRF2 greatly increases the rate of Holliday junction (HJ) formation and blocks the cleavage by various types of HJ resolving activities, including the newly identified human GEN1 protein. By using potassium permanganate probing and differential scanning calorimetry, we reveal that the basic domain of TRF2 induces structural changes to the junction. We propose that TRF2 contributes to t‐loop stabilisation by stimulating HJ formation and by preventing resolvase cleavage. These findings provide novel insights into the interplay between telomere protection and homologous recombination and suggest a general model in which TRF2 maintains telomere integrity by controlling the turnover of HJ at t‐loops and at regressed replication forks.

Molten salts database for energy applications

Abstract The growing interest in energy applications of molten salts is justified by several of their properties. Their possibilities of usage as a coolant, heat transfer fluid or heat storage substrate, require thermo-hydrodynamic refined calculations. Many researchers are using simulation techniques, such as Computational Fluid Dynamics (CFD) for their projects or conceptual designs. The aim of this work is providing a review of basic properties (density, viscosity, thermal conductivity and heat capacity) of the most common and referred salt mixtures. After checking data, tabulated and graphical outputs are given in order to offer the most suitable available values to be used as input parameters for other calculations or simulations. The reviewed values show a general scattering in characterization, mainly in thermal properties. This disagreement suggests that, in several cases, new studies must be started (and even new measurement techniques should be developed) to obtain accurate values.

Nonlinear analysis of the dynamics of DNA breathing.

The base pairs that encode the genetic information in DNA show large amplitude localized excitations called DNA breathing. We discuss the experimental observations of this phenomenon and its theoretical analysis. Starting from a model introduced to study the thermal denaturation of DNA, we show that it can qualitatively describe DNA breathing but is quantitatively not satisfactory. We show how the model can be modified to be quantitatively correct. This defines a nonlinear lattice model, which is interesting in itself because it has nonlinear localized excitations, forming a new class of discrete breather.

Modelling DNA at the mesoscale : a challenge for nonlinear science?

A vehicle which is capable of recognizing shapes in a predetermined area, comprising: a plurality of ultrasonic sensors, an encoder, map drawing means for sequentially and continuously drawing a map of the prescribed area determined by information received from the encoder and ultrasonic sensors, memory means for storing the map drawn by the map drawing means and control means for instructing rectilinear movement, starting stopping and turning of the vehicle so as to move in a serpentine fashion, wherein the ultrasonic sensors, encoder and map drawing means are operated by the control means, the memory means writes and stores a history of its own movements in the area and as information is received remembers detected information from the encoder, and a change of direction of the vehicle which is instructed by the control means is determined by information on the map as well as the areas through which the vehicle has previously passed.

Thermal denaturation of DNA studied with neutron scattering.

The melting transition of DNA, whereby the strands of the double-helix structure completely separate at a certain temperature, has been characterized using neutron scattering. A Bragg peak from B-form fiber DNA has been measured as a function of temperature, and its widths and integrated intensities have been interpreted using the Peyrard-Bishop-Dauxois model with only one free parameter. The experiment is unique, as it gives spatial correlation along the molecule through the melting transition where other techniques cannot.

Experimental and theoretical studies of sequence effects on the fluctuation and melting of short DNA molecules

Understanding the melting of short DNA sequences probes DNA at the scale of the genetic code and raises questions which are very different from those posed by very long sequences, which have been extensively studied. We investigate this problem by combining experiments and theory. A new experimental method allows us to make a mapping of the opening of the guanines along the sequence as a function of temperature. The results indicate that non-local effects may be important in DNA because an AT-rich region is able to influence the opening of a base pair which is about 10 base pairs away. An earlier mesoscopic model of DNA is modified to correctly describe the timescales associated with the opening of individual base pairs well below melting, and to properly take into account the sequence. Using this model to analyze some characteristic sequences for which detailed experimental data on the melting is available (Montrichok et al 2003 Europhys. Lett. 62 452), we show that we have to introduce non-local effects of AT-rich regions to get acceptable results. This brings a second indication that the influence of these highly fluctuating regions of DNA on their neighborhood can extend to some distance.

Thermodynamics of protein‐cation interaction: Ca+2 and Mg+2 binding to the fifth binding module of the LDL receptor

The ligand binding domain of the LDL receptor (LDLR) contains seven structurally homologous repeats. The fifth repeat (LR5) is considered to be the main module responsible for the binding of lipoproteins LDL and β‐VLDL. LR5, like the other homologous repeats, is around 40‐residue long and contains three disulfide bonds and a conserved cluster of negatively charged residues surrounding a hexacoordinated calcium ion. The calcium coordinating cage is formed by the backbone oxygens of W193 and D198, and side‐chain atoms of D196, D200, D206, and E207. The functionality of LDLR is closely associated with the presence of calcium. Magnesium ions are to some extent similar to calcium ions. However, they appear to be involved in different physiological events and their concentrations in extracellular and intracellular compartments are regulated by different mechanisms. Whether magnesium ions can play a role in the complex cycle of LDLR internalization and recycling is not known. We report here a detailed study of the interaction between LR5 and these two cations combining ITC, emission fluorescence, high resolution NMR, and MD simulations, at extracellular and endosomal pHs. Our results indicate that the conformational stability and internal dynamics of LR5 are strongly modulated by the specific bound cation. It appears that the difference in binding affinity for these cations is somewhat compensated by their different concentrations in late LDL‐associated endosomes. While the mildly acidic and calcium‐depleted environment in late endosomes has been proposed to contribute significantly to LDL release, the presence of magnesium might assist in efficient LDLR recycling. Proteins 2010.

Guanine radical chemistry reveals the effect of thermal fluctuations in gene promoter regions

DNA is not the static entity that structural pictures suggest. It has been longly known that it ‘breathes’ and fluctuates by local opening of the bases. Here we show that the effect of structural fluctuations, exhibited by AT-rich low stability regions present in some common transcription initiation regions, influences the properties of DNA in a distant range of at least 10 bp. This observation is confirmed by experiments on genuine gene promoter regions of DNA. The spatial correlations revealed by these experiments throw a new light on the physics of DNA and could have biological implications, for instance by contributing to the cooperative effects needed to assemble the molecular machinery that forms the transcription complex.

Adding a new dimension to DNA melting curves

Standard DNA melting curves record the separation of the two strands versus temperature, but they do not provide any information on the location of the opening. We introduce an experimental method which adds a new dimension to the melting curves of short DNA sequences by allowing us to record the degree of opening in several positions along the molecule all at once. This adds the spatial dimension to the melting curves and allows a precise investigation of the role of the base-pair sequence on the fluctuations and denaturation of the DNA double helix. We illustrate the power of the method by investigating the influence of an AT rich region on the fluctuations of neighboring domains.

Ab initio molecular dynamics: Relationship between structural phases and the sound velocity in dense hydrogen

The phase diagram and the possible stable structures of molecular solid hydrogen are intriguing physical phenomena that still remain to be fully unveiled. Particularly, its transition to metallic hydrogen at high pressures is currently a hot topic of discussion. This letter reports a simulation method that links the ab initio, quantum molecular dynamic and mechanical properties calculations to study the relation between the structural phase transitions and sound velocity in solid molecular hydrogen. The pressure range studied is from 0.1 GPa to 180 GPa, at 15 K temperature, thereby our aim is to simulate the conditions of manufacture, handling and early stages of compression of the target fuel used in confinement inertial fusion. Phase I degeneration below 1 GPa is discussed.