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

Numerical identification of a nonlinear diffusion law via regularization in Hilbert scales

We consider the reconstruction of a diffusion coefficient in a quasilinear elliptic problem from a single measurement of overspecified Neumann and Dirichlet data. The uniqueness for this parameter identification problem has been established by Cannon and we therefore focus on the stable solution in the presence of data noise. For this, we utilize a reformulation of the inverse problem as a linear ill-posed operator equation with perturbed data and operators. We are able to explicitly characterize the mapping properties of the corresponding operators which allow us to apply regularization in Hilbert scales. We can then prove convergence and convergence rates of the regularized reconstructions under very mild assumptions on the exact parameter. These are, in fact, already needed for the analysis of the forward problem and no additional source conditions are required. Numerical tests are presented to illustrate the theoretical statements.Comment: 17 pages, 2 figure