Drosophila SWR1 and NuA4 complexes are defined by DOMINO isoforms

Histone acetylation and deposition of H2A.Z variant are integral aspects of active transcription. In Drosophila, the single DOMINO chromatin regulator complex is thought to combine both activities via an unknown mechanism. Here we show that alternative isoforms of the DOMINO nucleosome remodeling ATPase, DOM-A and DOM-B, directly specify two distinct multi-subunit complexes. Both complexes are necessary for transcriptional regulation but through different mechanisms. The DOM-B complex incorporates H2A.V (the fly ortholog of H2A.Z) genome-wide in an ATP-dependent manner, like the yeast SWR1 complex. The DOM-A complex, instead, functions as an ATP-independent histone acetyltransferase complex similar to the yeast NuA4, targeting lysine 12 of histone H4. Our work provides an instructive example of how different evolutionary strategies lead to similar functional separation. In yeast and humans, nucleosome remodeling and histone acetyltransferase complexes originate from gene duplication and paralog specification. Drosophila generates the same diversity by alternative splicing of a single gene

Kreuth V initiative: European consensus proposals for treatment of hemophilia using standard products, extended half-life coagulation factor concentrates and non-replacement therapies

This report contains the updated consensus recommendations for optimal hemophilia care produced in 2019 by three Working Groups (WG) on behalf of the European Directorate for Quality of Medicines and Healthcare in the frame of the Kreuth V Initiative. WG1 recommended access to prophylaxis for all patients, the achievement of plasma factor trough levels of at least 3-5% when extended half-life factor VIII (FVIII) and FIX products are used, a personalized treatment regimen, and a choice of chromogenic assays for treatment monitoring. It was also emphasized that innovative therapies should be supervised by hemophilia comprehensive care centers. WG2 recommended mandatory collection of postmarketing data to assure the long-term safety and efficacy of new hemophilia therapies, the establishment of national patient registries including the core data recommended by the European Medicines Agency and the International Society on Thrombosis and Haemostasis, with adequate support under public control, and greater collaboration to facilitate a comprehensive data evaluation throughout Europe. WG3 discussed methodological aspects of hemophilia care in the context of access decisions, particularly for innovative therapies, and recommended that clinical studies should be designed to provide the quality of evidence needed by regulatory authorities, HTA bodies and healthcare providers. The dialogue between all stakeholders in hemophilia care and patient organizations should be fostered to implement these recommendations

Unscheduled DNA replication in G1 causes genome instability and damage signatures indicative of replication collisions

DNA replicates once per cell cycle. Interfering with the regulation of DNA replication initiation generates genome instability through over-replication and has been linked to early stages of cancer development. Here, we engineer genetic systems in budding yeast to induce unscheduled replication in a G1- like cell cycle state. Unscheduled G1 replication initiates at canonical S-phase origins. We quantifiy the composition of replisomes in G1- and S-phase and identified firing factors, polymerase α, and histone supply as factors that limit replication outside S-phase. G1 replication per se does not trigger cellular checkpoints. Subsequent replication during S-phase, however, results in overreplication and leads to chromosome breaks and chromosome-wide, strandbiased occurrence of RPA-bound single-stranded DNA, indicating head-to-tail replication collisions as a key mechanism generating genome instability upon G1 replication. Low-level, sporadic induction of G1 replication induces an identical response, indicating findings from synthetic systems are applicable to naturally occurring scenarios of unscheduled replication initiation

Defining the RBPome of primary T helper cells to elucidate higher-order Roquin-mediated mRNA regulation

Post-transcriptional gene regulation in T cells is dynamic and complex as targeted transcripts respond to various factors. This is evident for the Icos mRNA encoding an essential costimulatory receptor that is regulated by several RNA-binding proteins (RBP), including Roquin-1 and Roquin-2. Here, we identify a core RBPome of 798 mouse and 801 human T cell proteins by utilizing global RNA interactome capture (RNA-IC) and orthogonal organic phase separation (OOPS). The RBPome includes Stat1, Stat4 and Vav1 proteins suggesting unexpected functions for these transcription factors and signal transducers. Based on proximity to Roquin-1, we select \~50 RBPs for testing coregulation of Roquin-1/2 targets by induced expression in wild-type or Roquin-1/2-deficient T cells. Besides Roquin-independent contributions from Rbms1 and Cpeb4 we also show Roquin-1/2-dependent and target-specific coregulation of Icos by Celf1 and Igf2bp3. Connecting the cellular RBPome in a post-transcriptional context, we find contributions from multiple RBPs to the prototypic regulation of mRNA targets by individual trans-acting factors

Transcription Factor MAFF (MAF Basic Leucine Zipper Transcription Factor F) Regulates an Atherosclerosis Relevant Network Connecting Inflammation and Cholesterol Metabolism

BACKGROUND: Coronary artery disease (CAD) is a multifactorial condition with both genetic and exogenous causes. The contribution of tissue-specific functional networks to the development of atherosclerosis remains largely unclear. The aim of this study was to identify and characterize central regulators and networks leading to atherosclerosis. METHODS: Based on several hundred genes known to affect atherosclerosis risk in mouse (as demonstrated in knockout models) and human (as shown by genome-wide association studies), liver gene regulatory networks were modeled. The hierarchical order and regulatory directions of genes within the network were based on Bayesian prediction models, as well as experimental studies including chromatin immunoprecipitation DNA-sequencing, chromatin immunoprecipitation mass spectrometry, overexpression, small interfering RNA knockdown in mouse and human liver cells, and knockout mouse experiments. Bioinformatics and correlation analyses were used to clarify associations between central genes and CAD phenotypes in both human and mouse. RESULTS: The transcription factor MAFF (MAF basic leucine zipper transcription factor F) interacted as a key driver of a liver network with 3 human genes at CAD genome-wide association studies loci and 11 atherosclerotic murine genes. Most importantly, expression levels of the low-density lipoprotein receptor (LDLR) gene correlated with MAFF in 600 CAD patients undergoing bypass surgery (STARNET [Stockholm-Tartu Atherosclerosis Reverse Network Engineering Task]) and a hybrid mouse diversity panel involving 105 different inbred mouse strains. Molecular mechanisms of MAFF were tested in noninflammatory conditions and showed positive correlation between MAFF and LDLR in vitro and in vivo. Interestingly, after lipopolysaccharide stimulation (inflammatory conditions), an inverse correlation between MAFF and LDLR in vitro and in vivo was observed. Chromatin immunoprecipitation mass spectrometry revealed that the human CAD genome-wide association studies candidate BACH1 (BTB domain and CNC homolog 1) assists MAFF in the presence of lipopolysaccharide stimulation with respective heterodimers binding at the MAF recognition element of the LDLR promoter to transcriptionally downregulate LDLR expression. CONCLUSIONS: The transcription factor MAFF was identified as a novel central regulator of an atherosclerosis/CAD-relevant liver network. MAFF triggered context-specific expression of LDLR and other genes known to affect CAD risk. Our results suggest that MAFF is a missing link between inflammation, lipid and lipoprotein metabolism, and a possible treatment target

Energy Frontier Research Center for Solid-State Lighting Science: Exploring New Materials Architectures and Light Emission Phenomena

The Energy Frontier Research Center (EFRC) for Solid-State Lighting Science (SSLS) is one of 46 EFRCs initiated in 2009 to conduct basic and use-inspired research relevant to energy technologies. The overarching theme of the SSLS EFRC is the exploration of energy conversion in tailored photonic structures. In this article we review highlights from the research of the SSLS EFRC. Major research themes include: studies of the materials properties and emission characteristics of III-nitride semiconductor nanowires; development of new phosphors and II−VI quantum dots for use as wavelength downconverters; fundamental understanding of competing radiative and nonradiative processes in current-generation, planar light-emitting diode architectures; understanding of the electrical, optical, and structural properties of defects in InGaN materials and heterostructures; exploring ways to enhance spontaneous emission through modification of the environment in which the emission takes place; and investigating routes such as stimulated emission that might outcompete nonradiative processes

Role of PLK1 in steroid hormone signaling in breast cancer cells

Steroid receptors (SRs) are hormone-induced transcription factors that regulate gene expression by their concerted actions with coregulatory proteins. Performing a quantitative mass spectrometry-based interaction screen in breast cancer cells, we identified Polo-like kinase 1 to interact with progesterone receptor (PR) and estrogen receptor (ER) in presence of their respective hormone stimuli. PLK1 is recruited to chromatin and regulates SR-dependent and –independent gene transcription. Global gene expression analysis let us observe that PLK1-coactivated genes are enriched in developmental functions and linked to good clinical prognosis in breast cancer patients. Using large-scale phosphoproteomics with MCF7 breast cancer cells treated with estradiol in presence or absence of the specific PLK1 inhibitor BI2536, we observed that 28% of estradiol-induced phosphorylation sites depended on PLK1 activity. Among proteins with PLK1-dependent phosphorylation sites, we identified several transcriptional regulators including the ER-associated histone H3K4-methylase MLL2. As PLK1 inhibition reduced the estradiol-mediated induction of H3K4 methylation levels at MLL2 target genes, together these results propose a role of PLK1 in the regulation of MLL2 activity. Finally, in a targeted approach using recombinant histone proteins as substrate of PLK1, we observed a specific phosphorylation of histone H2B at serine 36. In summary, we conclude that PLK1 affects SR-dependent and SR-independent gene transcription by phosphorylating several transcriptional regulators and chromatin components

Kriterien für die Akzeptanz von recyceltem Phosphatdünger aus Abwasser und Klärschlamm im ökologischen Landbau

Plant available phosphorus is a limiting factor in organic farming. In the wake of an increasing awareness of the planetary boundaries of phosphate rock deposits, various research efforts to develop fertilizers based on recycled phosphorus from wastewater, sewage sludge and sewage sludge ashes have been initiated. The study aimed at the future potential of these fertilizers for organic farming and tried to identify and evaluate criteria that need to be fulfilled so that fertilizers of recycled phosphorus would be accepted by organic farmers. The survey revealed that the most important criterion for the acceptance is the non-existence of hazardous substances in the fertilizer. Plant availability, low energy input and high transparency of the production process of the fertilizers were also regarded as relatively important. Spreadability, price, public acceptance and superficial distinction compared to mineral fertilizers play a minor role for the acceptance

Functional and Radiological Outcomes after Treatment with Custom-Made Glenoid Components in Revision Reverse Shoulder Arthroplasty

Glenoid implant position and fixation are challenging in severe glenoid defects in reverse total shoulder arthroplasty (rTSA). Custom-made glenoid implants are metal augmented implants that are specially produced for a certain defect. They provide the restoration of the joint line and proper fixation. This retrospective data analysis investigated the clinical and radiological outcomes after revision using custom-made glenoid implants. Between 2018 and 2020, nine patients (10 shoulders) with severe glenoid defects underwent revision rTSA using a custom-made glenoid implant (Materialise Glenius or Lima ProMade). The pre- and postoperative Constant Murley Score (CMS), UCLA Score and Subjective Shoulder Value (SSV) were assessed. Postoperative CT scans and X-rays in two planes were available. The minimum follow-up was 12 months, with a mean follow-up of 23.1 months. The mean preoperative CMS, UCLA Score and SSV were 10.9, 4.1 and 11.0, respectively. The mean postoperative CMS, UCLA Score and SSV showed significant increases of 51.7 (<0.001), 22.9 (<0.001) and 52.0 (<0.001), respectively. There were no signs of loosening implants or scapular notching, and no revision was necessary. This trial showed promising clinical and radiological short-term outcomes for custom-made glenoid components in revision rTSA