Structure of mammalian AMPK and its regulation by ADP

The heterotrimeric AMP-activated protein kinase (AMPK) has a key role in regulating cellular energy metabolism; in response to a fall in intracellular ATP levels it activates energy-producing pathways and inhibits energy-consuming processes. AMPK has been implicated in a number of diseases related to energy metabolism including type 2 diabetes, obesity and, most recently, cancer. AMPK is converted from an inactive form to a catalytically competent form by phosphorylation of the activation loop within the kinase domain: AMP binding to the γ-regulatory domain promotes phosphorylation by the upstream kinase, protects the enzyme against dephosphorylation, as well as causing allosteric activation. Here we show that ADP binding to just one of the two exchangeable AXP (AMP/ADP/ATP) binding sites on the regulatory domain protects the enzyme from dephosphorylation, although it does not lead to allosteric activation. Our studies show that active mammalian AMPK displays significantly tighter binding to ADP than to Mg-ATP, explaining how the enzyme is regulated under physiological conditions where the concentration of Mg-ATP is higher than that of ADP and much higher than that of AMP. We have determined the crystal structure of an active AMPK complex. The structure shows how the activation loop of the kinase domain is stabilized by the regulatory domain and how the kinase linker region interacts with the regulatory nucleotide-binding site that mediates protection against dephosphorylation. From our biochemical and structural data we develop a model for how the energy status of a cell regulates AMPK activity.     

Short-term international migration trends in England and Wales from 2004 to 2009

Short-term migration estimates for England and Wales are the latest addition to the Office for National Statistics (ONS) migration statistics. This article discusses definitions of short-term migration and the methodology that is used to produce the estimates. Some of the estimates and the changes in the estimates over time are then discussed. The article includes previously unpublished short-term migration statistics and therefore helps to give a more complete picture of the size and characteristics of short-term international migration for England and Wales than has previously been possible. ONS have identified a clear user requirement for short-term migration estimates at local authority (LA) level. Consequently, attention is also paid to the progress that has been made and future work that is planned to distribute England and Wales short-term migration estimates to LA level.     

The Drosophila melanogaster Genetic Reference Panel

A major challenge of biology is understanding the relationship between molecular genetic variation and variation in quantitative traits, including fitness. This relationship determines our ability to predict phenotypes from genotypes and to understand how evolutionary forces shape variation within and between species. Previous efforts to dissect the genotype-phenotype map were based on incomplete genotypic information. Here, we describe the Drosophila melanogaster Genetic Reference Panel (DGRP), a community resource for analysis of population genomics and quantitative traits. The DGRP consists of fully sequenced inbred lines derived from a natural population. Population genomic analyses reveal reduced polymorphism in centromeric autosomal regions and the X chromosome, evidence for positive and negative selection, and rapid evolution of the X chromosome. Many variants in novel genes, most at low frequency, are associated with quantitative traits and explain a large fraction of the phenotypic variance. The DGRP facilitates genotype-phenotype mapping using the power of Drosophila genetics.     

Satisfaction Experiences with Tenancy Mediation: Why is it so Successful?

Tenancy mediation is a form of systematic practice that has become firmly established in a number of different legislatures. Yet there has been little research on what makes this an effective or satisfactory procedure. This paper presents a study based on quantitative and qualitative research that looks at tenancy mediation in New Zealand. Drawing on the method of appreciative inquiry, it finds that a consistently high rate of participant satisfaction with voluntary mediation comes from three main components of mediation practice. These are to do with three-way interactive mediator assistance, recognition and reframing, and ‘trust in practice.’ Together, these often generate an experience of transformative mediation which underpins the satisfaction that disputants report.     

The business team standard: A means of improving the effectiveness of individual businesses in a multibusiness corporation

The paper describes a project carried out within a major chemicals corporation to improve the performance of the individual businesses. This was to be done by clarifying some of the organisational uncertainties in its structure and improving the way that specialists form coalitions to address market challenges together. The approach used was based on “systems thinking”, which is an intellectual framework of knowledge that attempts to view organisations as wholes and which studies the processes of change in any part in the context of the whole organisation. Some of the important concepts of systems thinking are explored as they might be applied within a business organisation. Specifically the tool used was the Viable System Model of Stafford Beer, which the authors interpreted and developed into a set of statements (“a Standard”) which describe best practice in such organisations. Managers have used this to explore possible gaps in their organisations and, with this understanding, find ways to improve performance.     

Monovalent cation leaks in human red cells caused by single amino-acid substitutions in the transport domain of the band 3 chloride-bicarbonate exchanger, AE1

We identified 11 human pedigrees with dominantly inherited hemolytic anemias in both the hereditary stomatocytosis and spherocytosis classes. Affected individuals in these families had an increase in membrane permeability to Na and K that is particularly marked at 0 degrees C. We found that disease in these pedigrees was associated with a series of single amino-acid substitutions in the intramembrane domain of the erythrocyte band 3 anion exchanger, AE1. Anion movements were reduced in the abnormal red cells. The 'leak' cation fluxes were inhibited by SITS, dipyridamole and NS1652, chemically diverse inhibitors of band 3. Expression of the mutated genes in Xenopus laevis oocytes induced abnormal Na and K fluxes in the oocytes, and the induced Cl transport was low. These data are consistent with the suggestion that the substitutions convert the protein from an anion exchanger into an unregulated cation channel.     

A common polymorphism acts as an intragenic modifier of mutant p53 behaviour

The p73 protein, a homologue of the tumour-suppressor protein p53, can activate p53-responsive promoters and induce apoptosis in p53-deficient cells. Here we report that some tumour-derived p53 mutants can bind to and inactivate p73. The binding of such mutants is influenced by whether TP53 (encoding p53) codon 72, by virtue of a common polymorphism in the human population, encodes Arg or Pro. The ability of mutant p53 to bind p73, neutralize p73-induced apoptosis and transform cells in cooperation with EJ-Ras was enhanced when codon 72 encoded Arg. We found that the Arg-containing allele was preferentially mutated and retained in squamous cell tumours arising in Arg/Pro germline heterozygotes. Thus, inactivation of p53 family members may contribute to the biological properties of a subset of p53 mutants, and a polymorphic residue within p53 affects mutant behaviour.