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

A component-based middleware framework for configurable and reconfigurable Grid computing

Significant progress has been made in the design and development of Grid middleware which, in its present form, is founded on Web services technologies. However, we argue that present-day Grid middleware is severely limited in supporting projected next-generation applications which will involve pervasive and heterogeneous networked infrastructures, and advanced services such as collaborative distributed visualization. In this paper we discuss a new Grid middleware framework that features (i) support for advanced network services based on the novel concept of pluggable overlay networks, (ii) an architectural framework for constructing bespoke Grid middleware platforms in terms of 'middleware domains' such as extensible interaction types and resource discovery. We believe that such features will become increasingly essential with the emergence of next-generation e-Science applications. Copyright (c) 2005 John Wiley & Sons, Ltd

Careers Service Work with Adults: A Survey

This report summarises the findings from a postal survey of all Careers Service Companies in England about their adult guidance services and services to employers. The survey was conducted by the Centre for Guidance Studies (University of Derby) and the National Institute for Careers Education and Counselling (NICEC) for the Department for Education and Employment (DfEE)

Guidance and Individual Learning Accounts

This report presents the results of a CeGS/NICEC project on the role of formal and informal guidance in support of Individual Learning Accounts. It draws on five casestudies of good/interesting practice in implementing this role, including learner profiles, and supported by a literature review. It concludes that guidance can play a significant role in engaging non-traditional learners; that it needs to be marketed more effectively; that a strategy is needed for funding for in-depth guidance; and that the use of ILAs themselves as a route for such funding should be encouraged

Guidance and Individual Learning Accounts

This report presents the results of a CeGS/NICEC project on the role of formal and informal guidance in support of Individual Learning Accounts. It draws on five casestudies of good/interesting practice in implementing this role, including learner profiles, and supported by a literature review. It concludes that guidance can play a significant role in engaging non-traditional learners; that it needs to be marketed more effectively; that a strategy is needed for funding for in-depth guidance; and that the use of ILAs themselves as a route for such funding should be encouraged