21,317 research outputs found

    The Eastward Enlargement of the Eurozone: Trade and FDI

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    Trade and FDI, Economic Integration

    Improved Constraints on Cosmic Microwave Background Secondary Anisotropies from the Complete 2008 South Pole Telescope Data

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    We report measurements of the cosmic microwave background (CMB) power spectrum from the complete 2008 South Pole Telescope (SPT) data set. We analyze twice as much data as the first SPT power spectrum analysis, using an improved cosmological parameter estimator which fits multi-frequency models to the SPT 150 and 220 GHz bandpowers. We find an excellent fit to the measured bandpowers with a model that includes lensed primary CMB anisotropy, secondary thermal (tSZ) and kinetic (kSZ) Sunyaev-Zel'dovich anisotropies, unclustered synchrotron point sources, and clustered dusty point sources. In addition to measuring the power spectrum of dusty galaxies at high signal-to-noise, the data primarily constrain a linear combination of the kSZ and tSZ anisotropy contributions at 150 GHz and ℓ = 3000: D^(tSZ) ^(3000) + 0.5 D_(kSZ)^(3000) = 4.5 ± 1.0 ÎŒK^2. The 95% confidence upper limits on secondary anisotropy power are D ^(tSZ)_(3000) < 5.3 ÎŒK^2 and D^(kSZ)_(3000) < 6.5 ÎŒK^2. We also consider the potential correlation of dusty and tSZ sources and find it incapable of relaxing the tSZ upper limit. These results increase the significance of the lower than expected tSZ amplitude previously determined from SPT power spectrum measurements. We find that models including non-thermal pressure support in groups and clusters predict tSZ power in better agreement with the SPT data. Combining the tSZ power measurement with primary CMB data halves the statistical uncertainty on σ8. However, the preferred value of σ8 varies significantly between tSZ models. Improved constraints on cosmological parameters from tSZ power spectrum measurements require continued progress in the modeling of the tSZ power

    Quantitative investigation of protein-RNA interactions and regulation by phosphorylation

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    Phosphorylierung modulieren. Obwohl heute bereits Tausende von Phosphorylierungsstellen annotiert sind, sind entsprechende funktionelle Informationen begrenzt. Dies ist zum Teil darauf zurĂŒckzufĂŒhren, dass es keine Hochdurchsatzmethoden zur Erforschung der Funktion einer Phosphorylierungsstelle gibt. Um dieser Herausforderung zu begegnen, habe ich eine auf Shotgun-Proteomik basierende Strategie zur Messung der RNA-BindungsaktivitĂ€t von RBPs und ihren phosphorylierten Proteoformen entwickelt, die 'quantitative RNA-Interactome Capture (qRIC)' genannt wird. QRIC quantifiziert die Pull-Down-Effizienz von RBPs, die mit Oligo(dT)-Magnetbeads isoliert werden. Diese Effizienz korreliert mit der Anzahl der RNA-Bindungsstellen und der SpezifitĂ€t der Motivbindung, und spiegelt so die RNA-Bindung in vivo wieder. In einer GegenĂŒberstellung der Pull-Down-Effizienz verschiedener Proteoformen in unbehandelten Zellen, habe ich qRIC als unvoreingenommenes Screening von regulatorischen Phosphorylierungsstellen in RBPs eingesetzt. FĂŒr jede einzelne Phosphorylierungsstelle wurde ein Delta-Effizienzwert berechnet, der den Einfluss auf die RNA-Bindung in vivo reflektiert. Die Effizienzunterschiede spiegelten das erwartete Verhalten von RBPs wĂ€hrend der Phasentrennung von membranlosen Organellen und die Ladungsabstoßung zwischen Phosphorylierungsstellen und Nukleotiden bei physiologischem pH-Wert wider. Mithilfe des Delta-Effizienzwertes identifizierte ich mehrere bereits bekannte regulatorische Phosphorylierungsstellen in SF3B1, UPF1 und ELAVL1, sowie neue, bisher unbekannte und möglicherweise regulatorische Phosphorylierungsstellen in SERBP1, LARP1 und RBM20. Phosphomimetische Mutationsvarianten dieser Phosphorylierungsstellen wurden analysiert, um den molekularen Einfluss auf die Regulation der RBP-Funktion zu untersuchen. Es konnte gezeigt werden, dass die Phosphorylierung bestimmter Stellen im Spleißregulator RBM20 dessen nukleo-zytoplasmatische Lokalisierung, die Assoziation mit zytosolischen RNA-Granula und die Spleißfunktion beeinflusst. Diese Erkenntnisse könnten sich beispielsweise auf die Entwicklung neuer Behandlungsmethoden fĂŒr Patienten mit dysfunktionalen RBM20-Mutationen auswirken, die zu dilatativer Kardiomyopathie fĂŒhren. QRIC kann als Hochdurchsatzverfahren dazu beitragen, unser Wissen ĂŒber die Regulierung von Protein-RNA-Interaktionen durch Phosphorylierung zu erweitern.Post-transcriptional regulation of gene expression is fundamental in health and disease. RNA-binding proteins (RBPs) directly bind and govern the fate of RNAs in cells. At the same time, cell signaling cascades control RBP functions by modulating their physicochemical properties through post-translational modifications, like phosphorylation. Although thousands of phosphorylation sites have been annotated, functional information is limited. This, in part, is due to the lack of high-throughput methods that measure function. To tackle this challenge I developed a shotgun proteomics-based strategy for measuring the RNA-binding activity of RBPs and their phosphorylated proteoforms, named quantitative RNA-interactome capture (qRIC). In qRIC, pull-down efficiency of RBPs isolation with oligo(dT) magnetic beads is quantified in cells at steady state and correlates with the number of RNA-binding sites and motif binding specificity, reflecting a link to RNA-binding in vivo. By contrasting pull-down efficiency of different proteoforms in the cells, I applied qRIC as an unbiased screening of regulatory phosphorylation sites in RBPs affecting pull-down efficiency. A delta efficiency score was calculated for each individual phosphorylation site to denote its influence on RNA-binding in vivo. Efficiency differences globally reflected the expected behavior of RBPs during phase separation of membraneless organelles and charge repulsion between phosphorylation sites and nucleotides in physiological pH. Using the delta efficiency score, I identified several previously known regulatory phosphorylation sites in SF3B1, UPF1 and ELAVL1, plus novel candidate regulatory sites in SERBP1, LARP1 and RBM20. Phosphomimetic mutant variants of these sites were analysed to investigate the molecular mechanism of regulation. Importantly, I show that phosphorylation of candidate sites in the splicing regulator RBM20 affects its nucleo-cytoplasmic localization, association with cytosolic RNA granules, and splicing function. These findings could have implications for the development of novel treatments based on kinase activity for patients with dysfunctional RBM20 mutations leading to congenital dilated cardiomyopathy. I anticipate that qRIC, as a high throughput approach, will help to expand our knowledge about the regulation of protein-RNA interactions and their regulation by phosphorylation
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