Functional and proteomic analysis of a full thickness filaggrin-deficient skin organoid model.

Atopic eczema is an itchy inflammatory disorder characterised by skin barrier dysfunction. Loss-of-function mutations in the gene encoding filaggrin ( ) are a major risk factor, but the mechanisms by which filaggrin haploinsufficiency leads to atopic inflammation remain incompletely understood. Skin as an organ that can be modelled using primary cells provides the opportunity for selected genetic effects to be investigated in detail.

EMSY expression affects multiple components of the skin barrier with relevance to atopic dermatitis.

Background: Atopic dermatitis (AD) is a common, complex, and highly heritable inflammatory skin disease. Genome-wide association studies offer opportunities to identify molecular targets for drug development. A risk locus on chromosome 11q13.

A domestication history of dynamic adaptation and genomic deterioration in Sorghum.

The evolution of domesticated cereals was a complex interaction of shifting selection pressures and repeated episodes of introgression. Genomes of archaeological crops have the potential to reveal these dynamics without being obscured by recent breeding or introgression. We report a temporal series of archaeogenomes of the crop sorghum (Sorghum bicolor) from a single locality in Egyptian Nubia.

The FANCD2-FANCI complex is recruited to DNA interstrand crosslinks before monoubiquitination of FANCD2.

The Fanconi anaemia (FA) pathway is important for the repair of DNA interstrand crosslinks (ICL). The FANCD2-FANCI complex is central to the pathway, and localizes to ICLs dependent on its monoubiquitination. It has remained elusive whether the complex is recruited before or after the critical monoubiquitination.

Growth of large and highly ordered 2D crystals of a K⁺ channel, structural role of lipidic environment.

2D crystallography has proven to be an excellent technique to determine the 3D structure of membrane proteins. Compared to 3D crystallography, it has the advantage of visualizing the protein in an environment closer to the native one. However, producing good 2D crystals is still a challenge and little statistical knowledge can be gained from literature.

An approach to automated acquisition of cryoEM images from lacey carbon grids.

An approach to automated acquisition of cryoEM image data from lacey carbon grids using the Leginon program is described. Automated liquid nitrogen top up of the specimen holder dewar was used as a step towards full automation, without operator intervention during the course of data collection. During cryoEM studies of actin labelled with myosin V, we have found it necessary to work with lacey grids rather than Quantifoil or C-flat grids due to interaction of myosin V with the support film.

Object detection by correlation coefficients using azimuthally averaged reference projections.

A method of computing correlation coefficients for object detection that takes advantage of using azimuthally averaged reference projections is described and compared with two alternative methods-computing a cross-correlation function or a local correlation coefficient versus the azimuthally averaged reference projections. Two examples of an application from structural biology involving the detection of projection views of biological macromolecules in electron micrographs are discussed. It is found that a novel approach to computing a local correlation coefficient versus azimuthally averaged reference projections, using a rotational correlation coefficient, outperforms using a cross-correlation function and a local correlation coefficient in object detection from simulated images with a range of levels of simulated additive noise.

Microtubule structure at 8 A resolution.

We have obtained a 3D reconstruction of intact microtubules, using cryoelectron microscopy and image processing, at a resolution of about 8 A, sufficient to resolve much of the secondary structure. The level of detail in the map allows docking of the tubulin structure previously determined by electron crystallography, with very strong constraints, providing several important insights not previously available through docking tubulin into lower-resolution maps. This work provides an improved picture of the interactions between adjacent protofilaments, which are responsible for microtubule stability, and also suggests that some structural features are different in microtubules from those in the zinc sheets with which the tubulin structure was determined.