Probing the (H3-H4)(2) histone tetramer structure using pulsed EPR spectroscopy combined with site-directed spin labelling

The (H3-H4)2 histone tetramer forms the central core of nucleosomes and, as such, plays a prominent role in assembly, disassembly and positioning of nucleosomes. Despite its fundamental role in chromatin, the tetramer has received little structural investigation. Here, through the use of pulsed electron-electron double resonance spectroscopy coupled with site-directed spin labelling, we survey the structure of the tetramer in solution. We find that tetramer is structurally more heterogeneous on its own than when sequestered in the octamer or nucleosome. In particular, while the central region including the H3-H3′ interface retains a structure similar to that observed in nucleosomes, other regions such as the H3 αN helix display increased structural heterogeneity. Flexibility of the H3 αN helix in the free tetramer also illustrates the potential for post-translational modifications to alter the structure of this region and mediate interactions with histone chaperones. The approach described here promises to prove a powerful system for investigating the structure of additional assemblies of histones with other important factors in chromatin assembly/fluidity

The histone chaperones Nap1 and Vps75 bind histones H3 and H4 in a tetrameric conformation

Histone chaperones physically interact with histones to direct proper assembly and disassembly of nucleosomes regulating diverse nuclear processes such as DNA replication, promoter remodeling, transcription elongation, DNA damage, and histone variant exchange. Currently, the best-characterized chaperone-histone interaction is that between the ubiquitous chaperone Asf1 and a dimer of H3 and H4. Nucleosome assembly proteins (Nap proteins) represent a distinct class of histone chaperone. Using pulsed electron double resonance (PELDOR) measurements and protein crosslinking, we show that two members of this class, Nap1 and Vps75, bind histones in the tetrameric conformation also observed when they are sequestered within the nucleosome. Furthermore, H3 and H4 trapped in their tetrameric state can be used as substrates in nucleosome assembly and chaperone-mediated lysine acetylation. This alternate mode of histone interaction provides a potential means of maintaining the integrity of the histone tetramer during cycles of nucleosome reassembly

Oppositions and Coalitions: James Gardiner and Saskatchewan Provincial Politics, 1929 to 1934

James Gardiner devint le premier chef de gouvernement en Saskatchewan à perdre le pouvoir alors que le parti libéral qu'il dirigeait détenait le plus grand nombre de sièges. Après sa défaite, en 1929, il mit un temps considérable à déterminer la meilleure ligne de conduite à suivre pour son parti : il devint chef de l'Opposition en choisissant de l'être sans s'allier à aucun autre des partis en cause.Au cours des années qui suivirent, toutefois, les circonstances furent telles qu'il dût, en deux occasions, considérer la possibilité d'une coalition avec M.J. Coldwell et ses partisans. De plus, en 1932, il fut également sollicité par le gouvernement de J.T.M. Anderson en vue d'une coalition qui aurait permis à trois libéraux, dont Gardiner, de siéger au Cabinet. Cependant, rien dans tout cela ne cadrait avec la conception qu'entretenait Gardiner du rôle de chef de l'Opposition. Pour lui, ce rôle sous-entendait, à la fois, la préservation du système parlementaire et le maintien de celui des partis.Cette politique s'avéra d'ailleurs d'une grande importance dans l'histoire des partis politiques en Saskatchewan et pour le sien en particulier. Le talent qu'il mit à y maintenir une organisation politique vigoureuse, et ce, en dépit du fait qu'il était démuni de toutes les ressources financières dévolues au parti au pouvoir, fit en sorte que, dès 1934, la coalition qui gardait Anderson au pouvoir fut non seulement défaite mais détruite

Base Flow Characteristics for Several Four-Clustered Rocket Configurations at Mach Numbers from 2.0 to 3.5

A generalized study of base flow phenomena has been conducted with four 500-pound-thrust JP-4 fuel-liquid-oxygen rocket motors installed in the base of a 12-inch-diameter cylindrical model. Data were obtained over a Mach number and nozzle pressure ratio range of 2.0 to 3.5 and 340 to 600, respectively. Base heat flux, gas temperature, and pressure were highest in the center of the cluster core and decreased in a radial direction. Although a maximum heat flux of 93 Btu per square foot per second was measured within the cluster core, peripheral heat fluxes were low, averaging about 5 Btu per square foot per second for all configurations. Generally base heat flux was found to be independent of Mach number over the range investigated. Base heat flux within the cluster core was decreased by increasing motor spacing, motor extension, a combination of increasing nozzle area ratio and decreasing exit angle and gimbaling the two side engines. Small amounts of nitrogen injected within the cluster core sharply reduced core heat flux

The spatial effect of protein deuteration on nitroxide spin-label relaxation:implications for EPR distance measurement

This work was supported by a Wellcome Trust Senior Fellowship (095062) to T.O.-H. The Authors would also like to acknowledge funding from The MRC – United Kingdom, Grant G1100021.Pulsed electron-electron double resonance (PELDOR) coupled with site-directed spin labeling is a powerful technique for the elucidation of protein or nucleic acid, macromolecular structure and interactions. The intrinsic high sensitivity of electron paramagnetic resonance enables measurement on small quantities of bio-macromolecules, however short relaxation times impose a limit on the sensitivity and size of distances that can be measured using this technique. The persistence of the electron spin-echo, in the PELDOR experiment, is one of the most crucial limitations to distance measurement. At a temperature of around 50 K one of the predominant factors affecting persistence of an echo, and as such, the sensitivity and measurable distance between spin labels, is the electron spin echo dephasing time (Tm). It has become normal practice to use deuterated solvents to extend Tm and recently it has been demonstrated that deuteration of the underlying protein significantly extends Tm. Here we examine the spatial effect of segmental deuteration of the underlying protein, and also explore the concentration and temperature dependence of highly deuterated systems.Publisher PDFPeer reviewe

Involuntary automorphisms of groups of odd order.

One of the most important and far reaching concepts in mathematics is that of a group. It is to this branch of mathematics that the work in this thesis belongs. An automorphism of a group is an expression of the symmetry of the group structure. If there is sufficient symmetry then it is to be expected that the structure will be restricted. The purpose of this thesis is to make a precise statement of this relationship for the case in which the symmetry is given by an invelutory automorphism

First Principles Theory of the Lattice Thermal Conductivity of Semiconductors

Thesis advisor: David A. BroidoUsing density functional perturbation theory and a full solution of the linearized phonon Boltzmann transport equation (BTE), a parameter-free theory of semiconductor thermal properties is developed. The approximations and shortcomings of previous approaches to thermal conductivity calculations are investigated. The use of empirical interatomic potentials in the BTE approach is shown to give poor agreement with measured values of thermal conductivity. By using the adiabatic bond charge model, the importance of accurate descriptions of phonon dispersions is highlighted. The extremely limited capacity of previous theoretical techniques in the realm of thermal conductivity prediction is highlighted; this is due to a dependence on adjustable parameters. Density functional perturbation theory is coupled with an iterative solution to the full Boltzmann transport equation creating a theoretical construct where thermal conductivity prediction becomes possible. Validation of the approach is demonstrated through the calculation of a range of thermal properties for a set of polar and non-polar semiconductors which are compared with measured values. The agreement between theory and measurement is very good, confirming the promise of the theoretical approach. Due to the significant computational effort required by the parameter-free calculations, new forms for room temperature relaxation time approximations are derived. The resulting forms produce thermal conductivity values in very good agreement with the ab initio data across a wide temperature range. It is therefore shown that accurate relaxation time approximations can be developed, fixing the adjustable parameters to the ab initio theory avoiding any comparison with measured data. This approach improves the accuracy of phonon relaxation times compared with previous models.Thesis (PhD) — Boston College, 2009.Submitted to: Boston College. Graduate School of Arts and Sciences.Discipline: Physics