Chromatin, the physiological template of all eukaryotic genetic information, is subject to a diverse array of posttranslational modifications that largely impinge on histone amino termini, thereby regulating access to the underlying DNA. Distinct histone amino-terminal modifications can generate synergistic or antagonistic interaction affinities for chromatin-associated proteins, which in turn dictate dynamic transitions between transcriptionally active or transcriptionally silent chromatin states. The combinatorial nature of histone amino-terminal modifications thus reveals a “histone code” that considerably extends the information potential of the genetic code. We propose that this epigenetic marking system represents a fundamental regulatory mechanism that has an impact on most, if not all, chromatin-templated processes, with far-reaching consequences for cell fate decisions and both normal and pathological development.

/pdf/translating-the-histone-code-l291mddcr9.pdf

Translating the Histone Code

Histone proteins and the nucleosomes they form with DNA are the fundamental building blocks of eukaryotic chromatin. A diverse array of post-translational modifications that often occur on tail domains of these proteins has been well documented. Although the function of these highly conserved modifications has remained elusive, converging biochemical and genetic evidence suggests functions in several chromatin-based processes. We propose that distinct histone modifications, on one or more tails, act sequentially or in combination to form a 'histone code' that is, read by other proteins to bring about distinct downstream events.

The language of covalent histone modifications.

The X-ray crystal structure of the nucleosome core particle of chromatin shows in atomic detail how the histone protein octamer is assembled and how 146 base pairs of DNA are organized into a superhelix around it. Both histone/histone and histone/DNA interactions depend on the histone fold domains and additional, well ordered structure elements extending from this motif. Histone amino-terminal tails pass over and between the gyres of the DNA superhelix to contact neighbouring particles. The lack of uniformity between multiple histone/DNA-binding sites causes the DNA to deviate from ideal superhelix geometry.

Crystal structure of the nucleosome core particle at 2.8 Å resolution

DNA Double-stranded Breaks Induce Histone H2AX Phosphorylation on Serine 139

BIOE 402. Medical Technology Assessment. 2 or 3 hours. Bioentrepreneur course. Assessment of medical technology in the context of commercialization. Objectives, competition, market share, funding, pricing, manufacturing, growth, and intellectual property; many issues unique to biomedical products. Course Information: 2 undergraduate hours. 3 graduate hours. Prerequisite(s): Junior standing or above and consent of the instructor.

“Bioinformatics” 특집을 내면서

Solvent Mediated Interactions in the Structure of the Nucleosome Core Particle at 1.9 Å Resolution

Publisher Summary This chapter describes the preparation method of nucleosome core particles (NCPs) from recombinant histones. The ability to make defined NCPs, or arrays of nucleosomes, from histone proteins expressed in bacteria has several advantages over previously used methods using histones isolated from natural sources. The chapter discusses protocols for the overexpression and purification of histones H2A, H2B, H3, and H4, both as full-length proteins and as corresponding trypsin-resistant globular domains. The chapter presents a method for the refolding and purification of a histone octamer from denatured recombinant histone proteins together with a protocol for the assembly and purification of a nucleosome core particle using 146 base pairs of DNA. The purity and homogeneity of the final core-particle preparation are assessed by a high-resolution gel shift assay. The chapter presents a flow chart that describes the procedures involved in the preparation of synthetic nucleosomes.

Preparation of nucleosome core particle from recombinant histones.

Publisher Summary The ability to prepare nucleosome core particles (NCPs), or nucleosomal arrays, from recombinant histone proteins and defined-sequence DNA has become a requirement in many projects that address the role of histone modifications, histone variants, or histone mutations in nucleosome and chromatin structure. The cloning strategies for the construction of plasmids containing multiple repeats of defined DNA sequences, and the subsequent large-scale isolation of defined sequence DNA for nucleosome reconstitution are described. This chapter also describes adapted procedures to prepare nucleosomes with histones from other species, and for the refolding and reconstitution of (H2A– H2B) dimers and (H3–H4) 2 tetramers. Methods to reconstitute nucleosomes from different histone subcomplexes are described. A flow chart for all procedures involved in the preparation of synthetic nucleosomes is also presented.

Reconstitution of Nucleosome Core Particles from Recombinant Histones and DNA

Chromatin compaction has profound implications for the regulation of transcription, replication and DNA repair. Changes in nucleosome structure and stability — due to the incorporation of variant histones and post-translational modifications of histones — affect chromatin compaction. Chromatin structures are not nearly as uniform as previously assumed, which should be taken into account in the context of specific biological functions.

/pdf/new-insights-into-nucleosome-and-chromatin-structure-an-10iq6o59fb.pdf

Karolin Luger

Papers

Crystal structure of the nucleosome core particle at 2.8 Å resolution

Solvent Mediated Interactions in the Structure of the Nucleosome Core Particle at 1.9 Å Resolution

Preparation of nucleosome core particle from recombinant histones.

Reconstitution of Nucleosome Core Particles from Recombinant Histones and DNA

New insights into nucleosome and chromatin structure: an ordered state or a disordered affair?