The Arabidopsis NOT4A E3 ligase promotes PGR3 expression and regulates chloroplast translation

Chloroplast function requires the coordinated action of nuclear- and chloroplast-derived proteins, including several hundred nuclear-encoded pentatricopeptide repeat (PPR) proteins that regulate plastid mRNA metabolism. Despite their large number and importance, regulatory mechanisms controlling PPR expression are poorly understood. Here we show that the Arabidopsis NOT4A ubiquitin-ligase positively regulates the expression of PROTON GRADIENT REGULATION 3 (PGR3), a PPR protein required for translating several thylakoid-localised photosynthetic components and ribosome subunits within chloroplasts. Loss of NOT4A function leads to a strong depletion of cytochrome b6f and NAD(P)H dehydrogenase (NDH) complexes, as well as plastid 30 S ribosomes, which reduces mRNA translation and photosynthetic capacity, causing pale-yellow and slow-growth phenotypes. Quantitative transcriptome and proteome analysis of the not4a mutant reveal it lacks PGR3 expression, and that its molecular defects resemble those of a pgr3 mutant. Furthermore, we show that normal plastid function is restored to not4a through transgenic PGR3 expression. Our work identifies NOT4A as crucial for ensuring robust photosynthetic function during development and stress-response, through promoting PGR3 production and chloroplast translatio

What does supervision help with? a survey of 315 social workers in the UK

What does social work supervision help with? There are many different models of supervision and an increasing amount of research. Much of this is concerned with the content of supervision and how supervisors (and supervisees) should behave — and these are important concerns. But even more important is the question of who or what supervision helps with. Supervision is widely considered to have many different functions but in the context of UK local authority social work, must ultimately prove itself as a method for helping people who use services. This article reports on a survey of 315 social workers from UK local authorities. Most reported that supervision helps primarily with management oversight and accountability. However, the small number of practitioners who received regular group supervision and those who received supervision more frequently said it helped with a much broader range of things

'We don't learn democracy, we live it!' : consulting the pupil voice in Scottish schools

As the education for citizenship agenda continues to impact on schools, there is a need to begin the discussion around examining the kind of initiatives that can push it forward. In Scotland the proposals should, it is argued, permeate the curriculum throughout the school. Yet there is the fear that the responsibility of all can become the responsibility of none. This paper examines, through case study research carried out by the authors, initiatives in schools designed to take forward the citizenship agenda in the light of children's rights. The first two relate to firstly the impact of pupil councils in primary schools and secondly the impact of discussing controversial issues in the primary classroom. The third outlines the impact on values and dispositions of developing more participatory, democratic practice in the classroom. The paper concludes by calling for both more initiatives of this type and more evaluation of their worth

Simulating supervision: how do managers respond to a crisis?

Supervision is fundamental to child and family social work practice, in England as elsewhere, yet there is little research regarding what managers and social workers do when they meet to discuss the families they are working with. Recent years have seen a growing interest in the use of simulated clients and Objective Structured Clinical Exams to help develop and evaluate the abilities of social workers and students. This paper describes a study of 30 simulated supervision sessions between English social work managers and an actor playing the role of a student social worker in need of support. The simulation concerns a referral regarding an incident of domestic abuse. During the simulations, managers typically asked closed questions to obtain more information before providing solutions for the supervisee in the form of advice and direction. There was little evidence of emotional support for the social worker, nor empathy with the family. Managers typically acted as expert problem-solvers. The implications of this are discussed in relation to current theoretical models of supervision for child and family social work and in relation to how Children’s Services responds to domestic abuse

Sorting Signals, N-Terminal Modifications and Abundance of the Chloroplast Proteome

Characterization of the chloroplast proteome is needed to understand the essential contribution of the chloroplast to plant growth and development. Here we present a large scale analysis by nanoLC-Q-TOF and nanoLC-LTQ-Orbitrap mass spectrometry (MS) of ten independent chloroplast preparations from Arabidopsis thaliana which unambiguously identified 1325 proteins. Novel proteins include various kinases and putative nucleotide binding proteins. Based on repeated and independent MS based protein identifications requiring multiple matched peptide sequences, as well as literature, 916 nuclear-encoded proteins were assigned with high confidence to the plastid, of which 86% had a predicted chloroplast transit peptide (cTP). The protein abundance of soluble stromal proteins was calculated from normalized spectral counts from LTQ-Obitrap analysis and was found to cover four orders of magnitude. Comparison to gel-based quantification demonstrates that ‘spectral counting’ can provide large scale protein quantification for Arabidopsis. This quantitative information was used to determine possible biases for protein targeting prediction by TargetP and also to understand the significance of protein contaminants. The abundance data for 550 stromal proteins was used to understand abundance of metabolic pathways and chloroplast processes. We highlight the abundance of 48 stromal proteins involved in post-translational proteome homeostasis (including aminopeptidases, proteases, deformylases, chaperones, protein sorting components) and discuss the biological implications. N-terminal modifications were identified for a subset of nuclear- and chloroplast-encoded proteins and a novel N-terminal acetylation motif was discovered. Analysis of cTPs and their cleavage sites of Arabidopsis chloroplast proteins, as well as their predicted rice homologues, identified new species-dependent features, which will facilitate improved subcellular localization prediction. No evidence was found for suggested targeting via the secretory system. This study provides the most comprehensive chloroplast proteome analysis to date and an expanded Plant Proteome Database (PPDB) in which all MS data are projected on identified gene models

Current status of the multinational Arabidopsis community

The multinational Arabidopsis research community is highly collaborative and over the past thirty years these activities have been documented by the Multinational Arabidopsis Steering Committee (MASC). Here, we (a) highlight recent research advances made with the reference plant Arabidopsis thaliana; (b) provide summaries from recent reports submitted by MASC subcommittees, projects and resources associated with MASC and from MASC country representatives; and (c) initiate a call for ideas and foci for the “fourth decadal roadmap,” which will advise and coordinate the global activities of the Arabidopsis research community

Phylogenomic analysis of the Chlamydomonas genome unmasks proteins potentially involved in photosynthetic function and regulation

Chlamydomonas reinhardtii, a unicellular green alga, has been exploited as a reference organism for identifying proteins and activities associated with the photosynthetic apparatus and the functioning of chloroplasts. Recently, the full genome sequence of Chlamydomonas was generated and a set of gene models, representing all genes on the genome, was developed. Using these gene models, and gene models developed for the genomes of other organisms, a phylogenomic, comparative analysis was performed to identify proteins encoded on the Chlamydomonas genome which were likely involved in chloroplast functions (or specifically associated with the green algal lineage); this set of proteins has been designated the GreenCut. Further analyses of those GreenCut proteins with uncharacterized functions and the generation of mutant strains aberrant for these proteins are beginning to unmask new layers of functionality/regulation that are integrated into the workings of the photosynthetic apparatus

Mutagenesis and Functional Studies with Succinate Dehydrogenase Inhibitors in the Wheat Pathogen Mycosphaerella graminicola

A range of novel carboxamide fungicides, inhibitors of the succinate dehydrogenase enzyme (SDH, EC 1.3.5.1) is currently being introduced to the crop protection market. The aim of this study was to explore the impact of structurally distinct carboxamides on target site resistance development and to assess possible impact on fitness