76 research outputs found
DS_DISC874313 – Supplemental material for Development of a High-Throughput Fluorescence Polarization Assay to Detect Inhibitors of the FAK–Paxillin Interaction
No full textSupplemental material, DS_DISC874313 for Development of a High-Throughput Fluorescence Polarization Assay to Detect Inhibitors of the FAK–Paxillin Interaction by Timothy Marlowe, Carlos Alvarado, Andrew Rivera, Felicia Lenzo, Rohini Nott, Dena Bondugji, Justin Montoya, Alana Hurley, Matt Kaplan, Andrew Capaldi and William Cance in SLAS Discovery</p
Naples et Pompeii nell'Encyclopédie
No full textThe entries “Naples”, “Herculanum”, “Pompeii” in the Encyclopédie ou Dictionnaire raisonné des sciences, des arts et des métiers
(1751-1772) are of different lengths and depths. These are entries that on the one hand report some of the effects of the archeolo gical
discoveries which amaze Europe during those years, while on the other they impose themselves as testimonies of some explanatory
techniques (and of some imperfections) which are typical of the ripest “encyclopedism”. Their author De Jaucourt carries out a d edicated
and significant work of construction and divulgation and he brings to the attention of the subscribers of the Encyclopédie the treasures
at the core of the Vesuvius land
Imaging mass cytometry pseudoimage data underlying the publication "Decoding mitochondrial heterogeneity in single muscle fibres by imaging mass cytometry"
No full text.zip archive containing IMC pseudoimage data underlying the publication below:Decoding mitochondrial heterogeneity in single muscle fibres by imaging mass cytometryCharlotte Warren, David McDonald, Roderick Capaldi, David Deehan, Robert W. Taylor, Andrew Filby, Doug M. Turnbull, Conor Lawless & Amy E. VincentScientific Reports volume 10, Article number: 15336 (2020)https://doi.org/10.1038/s41598-020-70885-3</p
RECONSTRUCTING THE S. CEREVISIAE GROWTH CONTROL NETWORK IN STRESS CONDITIONS
No full textRights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Downloaded 13-Sep-2016 12:15:21 Link to ite
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Estimating the Rate of FOXO1 Phosphorylation and Dephosphorylation Using Live Cell Imaging
No full textFoxO1 is a signaling transcription factor regulated by the growth factor/PI3K/AKT pathway. Phosphorylation of FOXO1 by the serine/threonine kinase AKT, sequesters FOXO1 in the cytoplasm by blocking the interaction of FOXO1’s nuclear localization signal (NLS) with nuclear transport receptors and promoting FOXO1 binding to the cytoplasmic 14-3-3 proteins. Dephosphorylation of FOXO1 by the phosphatase PP2A restores NLS function and leads to accumulation of FOXO1 in the nucleus. Here, we use fluorescently labeled FOXO1 to characterize its nuclear trafficking dynamics under conditions of AKT and PP2A inhibition in order to describe the relative cytoplasmic dephosphorylation rate by PP2A and relative nuclear phosphorylation rate by AKT on FOXO1 respectively. Measured results affirm previous data that indicates AKT is less active in the nucleus than the cytoplasm and suggests that FOXO1 may undergo rapid shuttling into and out of the nucleus even during AKT activation
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The Analysis of mRNP Granule Composition and Structure in Saccharomyces cerevisiae
No full textA recurring theme in biology is the aggregation of mRNA-protein complexes (mRNPs) into higher order assemblies. Often these complexes play important roles in the regulation of gene expression, but the function of the conserved cytoplasmic mRNP assemblies - P bodies and stress granules, is not known. It is believed that the misregulation of granule assembly is related to disorders like Amyotrophic Lateral Sclerosis and Frontotemporal Lobe Degeneration. Determining the complete composition of these granules may hold the key to understanding the function and mechanism of assembly of these granules. This work describes multiple approaches taken to identify new protein and mRNA components of P bodies and stress granules. New members of the P body and stress granule proteome reveal a role for these granules in diverse cellular processes including signal transduction, transcription and metabolism. Additionally, a new stress granule resident complex - the CCT complex, was also identified as a novel regulator of granule disassembly. This work also describes the first purification scheme for stress granules and presents a new system for in vitro study of stress granules. Together, the findings shed new light on the composition, function, structure and regulation of P bodies and stress granules in yeast
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Multilayered Regulation of TORC1 Signaling in Saccharomyces cerevisiae
No full textThe Target of Rapamycin Complex 1 (TORC1) is a master regulator of cellular growth in eukaryotes. Much insight has been gained into how amino acid and nitrogen levels regulate TORC1 through the escape from rapamycin-induced growth arrest complex (EGOC), and its regulators including the Seh1-associated complex (SEAC). However, other nutrient levels and environmental stresses also act on TORC1, and far less is known about how these signals are transmitted to the complex. In two projects presented here we investigate the osmotic stress signaling network acting on TORC1 as well as regulators of TORC1 agglomeration that act in glucose and nitrogen starvation conditions.
In the first investigation, we introduce a novel and reproducible high-throughput assay to screen for genes that affect TORC1 activity in stress conditions. We then use these methods to measure the expression of a TORC1 dependent ribosome biogenesis gene, NSR1, in ~4700 strains from the yeast knock-out library during osmotic stress. We show that 440 of these strains are not able to properly repress NSR1 transcription. The genes identified in the screen form a highly-connected network including 17 proteins that directly interact with TORC1. Secondary rapamycin-based assays performed on these strains allowed us to further characterize the network and show that more than 50 of the proteins act downstream of TORC1. The data derived from this work serve as a resource for our lab and others studying TORC1, and the assay itself is customizable and can be used to characterize any gene regulatory network.
In the second study, we sought to further our understanding of the movement of TORC1 from its position distributed across the surface of the vacuolar membrane to a single agglomerate (TORC1-body) in starvation conditions. Previous work suggested that the AMPK in yeast, Snf1, indirectly promoted the phosphorylation of the TORC1 component Kog1. This phosphorylation event sped up aggregation of the complex by ~20 fold. In order to identify other signaling proteins that regulate TORC1-body formation we performed a screen examining the impact that nearly all non-essential kinases and phosphatases in yeast, as well as selected proteins from the previous high-throughput network, have on TORC1 agglomeration. We identified 13 new regulators of TORC1 body formation, including the PI(3)P binding protein Pib2. We also examined the impact of EGOC deletions and mutants had on body formation and discovered that active EGOC was an inhibitor of TORC1 aggregation. Together, we show that seven of the new regulators likely act at or above the EGOC dependent inhibition of TORC1 body formation; while others act at a later step to assist in body formation.Release after 01/02/202
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Chaperone Limitations in Prion Curing
No full textAmyloid promotes a dramatic transition in protein conformation that perpetuates, giving rise to a broad variety of distinct phenotypes, ranging from pathological disorders to dynamic heritable traits. Amyloid has long been thought to be resistant to clearance by the proteostasis network, but increasing evidence is challenging this view. For example, heat shock disassembles yeast prion amyloids, revealing in vivo solubilization of these aggregates. However, the exact proteostatic niche that promotes amyloid clearance is largely unknown. We identified several environmental stresses leading to prion curing via the same mechanism as heat shock and further showed that a shared characteristic was the activation of the transcription factor heat shock factor 1 (Hsf1). Strikingly, artificial Hsf1 activation interfered with heat shock-mediated prion curing, presumably due to overexpression of a nucleotide exchange factor Sse1. Limiting Sse1, which decelerates the Hsp70 cycle, promoted chaperone loading on prion aggregates and enabled artificial Hsf1 activation to resolve prion aggregates; in contrast, it impaired resolution of stress-induced aggregates and cell growth at elevated temperature. Thus, our study demonstrates that the proteostasis network, fine-tuned for optimal dissolution of non-amyloid aggregates, can be reconfigured for solubilization of amyloid by modulating the Hsp70 cycle.Release after 12/16/202
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