mp A meteorite explodes on the Moon: Q&A with geophysicist Bruce Campbell By insider.si.edu Published On :: Thu, 30 May 2013 09:49:04 +0000 A fiery explosion on the surface of the Moon, visible to the naked eye, recently surprised NASA astronomers monitoring the Moon for meteorite strikes. Occurring […] The post A meteorite explodes on the Moon: Q&A with geophysicist Bruce Campbell appeared first on Smithsonian Insider. Full Article Q & A Research News Science & Nature Space meteorites
mp Q&A: Plastics expert Odile Madden on plastic debris in Alaskan waters By insider.si.edu Published On :: Wed, 10 Jul 2013 14:16:26 +0000 In June, Odile Madden, materials scientist at the Smithsonian’s Museum Conservation Institute, was a participant on a 6-day interdisciplinary expedition to a number of beaches […] The post Q&A: Plastics expert Odile Madden on plastic debris in Alaskan waters appeared first on Smithsonian Insider. Full Article Marine Science Q & A Research News Science & Nature conservation conservation biology endangered species mammals materials science Museum Conservation Institute pollution
mp Zoo scientists find sudden stream temperature changes boost hellbender immune systems By insider.si.edu Published On :: Tue, 10 Sep 2013 18:24:39 +0000 Hellbenders, aquatic salamanders from the eastern United States, are surprisingly good at dealing with unpredictable weather. In a recent study published in the Journal of […] The post Zoo scientists find sudden stream temperature changes boost hellbender immune systems appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature carbon dioxide climate change conservation conservation biology Smithsonian Conservation Biology Institute Smithsonian's National Zoo
mp Effects of human impact are long lasting for forests in Northeast U.S. By insider.si.edu Published On :: Wed, 18 Sep 2013 14:48:31 +0000 Grow fast, die young is not a lifestyle normally associated with trees. But in the forests of the Northeastern United States the red maple follows […] The post Effects of human impact are long lasting for forests in Northeast U.S. appeared first on Smithsonian Insider. Full Article Research News Science & Nature biodiversity birds carbon dioxide climate change conservation conservation biology insects Smithsonian Conservation Biology Institute
mp Funeral flowers in the Stone Age: Q&A with Smithsonian anthropologist Reuven Yeshurun By insider.si.edu Published On :: Mon, 23 Sep 2013 06:56:35 +0000 Roses, carnations and lilies are today among the most popular flowers people use to express sympathy and condolence. The simple act of sending flowers to […] The post Funeral flowers in the Stone Age: Q&A with Smithsonian anthropologist Reuven Yeshurun appeared first on Smithsonian Insider. Full Article Anthropology Dinosaurs & Fossils Q & A Research News Science & Nature archaeology National Museum of Natural History prehistoric
mp Package-delivering drones? Q&A with Roger Connor of the National Air and Space Museum By insider.si.edu Published On :: Mon, 09 Dec 2013 06:00:33 +0000 Santa Claus and his sleigh full of gifts has been upstaged early this holiday season by news of autonomous drones possibly delivering packages to your […] The post Package-delivering drones? Q&A with Roger Connor of the National Air and Space Museum appeared first on Smithsonian Insider. Full Article History & Culture Q & A Science & Nature aeronautics aviation National Air and Space Museum technology
mp Carabidae in the colony, seven new beetles that bunk with ants: Q&A with Terry Erwin By insider.si.edu Published On :: Mon, 13 Jan 2014 13:48:29 +0000 Ants dominate the earth’s ecosystems and many are voracious predators that use their mandibles and sheer numbers to pin down and tear apart most other […] The post Carabidae in the colony, seven new beetles that bunk with ants: Q&A with Terry Erwin appeared first on Smithsonian Insider. Full Article Animals Q & A Research News Science & Nature biodiversity conservation biology insects National Museum of Natural History new species
mp The secret formula to feeding 900 babies: Scientists uncover milk composition of naked mole-rat queens By insider.si.edu Published On :: Mon, 31 Mar 2014 22:31:30 +0000 Parents normally feel the need to provide well for their kids. For humans, that number of offspring is usually in the single digits, but a […] The post The secret formula to feeding 900 babies: Scientists uncover milk composition of naked mole-rat queens appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature bees conservation biology mammals Smithsonian Environmental Research Center Smithsonian's National Zoo
mp Commercial shipping lanes changed in Panama to save humpback whales By insider.si.edu Published On :: Wed, 28 May 2014 19:07:39 +0000 The Republic of Panama’s proposal to implement four Traffic Separation Schemes for commercial vessels entering and exiting the Panama Canal and ports was approved unanimously […] The post Commercial shipping lanes changed in Panama to save humpback whales appeared first on Smithsonian Insider. Full Article Marine Science Research News Science & Nature Colombia conservation conservation biology Ecuador endangered species South America Tropical Research Institute whales
mp Slimy invaders, giant snails: Q&A with mollusk expert Ellen Strong By insider.si.edu Published On :: Fri, 18 Jul 2014 19:14:58 +0000 It is related to one of the world’s most destructive invasive species, and possibly the slimiest. Thirty-five pounds of live giant West African snails (Archachatina […] The post Slimy invaders, giant snails: Q&A with mollusk expert Ellen Strong appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature Caribbean conservation biology extinction invasive species National Museum of Natural History
mp Smithsonian & SVF launch rare-breed livestock conservation partnership By insider.si.edu Published On :: Thu, 31 Jul 2014 12:54:04 +0000 The Smithsonian Conservation Biology Institute and the SVF Foundation have launched a new collaboration to strengthen rare and endangered livestock breed conservation through the preservation […] The post Smithsonian & SVF launch rare-breed livestock conservation partnership appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature conservation conservation biology endangered species mammals Smithsonian Conservation Biology Institute Smithsonian's National Zoo veterinary medicine
mp Simple tips to keep your backyard birds healthy this winter By insider.si.edu Published On :: Fri, 19 Sep 2014 13:58:58 +0000 What crunchy food did Americans spend $5.5 billion on last year—with sales that spiked before snow and ice storms? If you guessed birdseed then you […] The post Simple tips to keep your backyard birds healthy this winter appeared first on Smithsonian Insider. Full Article Science & Nature birds Migratory Bird Center Smithsonian's National Zoo
mp Smithsonian Symposium: The Anthropocene By insider.si.edu Published On :: Tue, 07 Oct 2014 13:48:54 +0000 Archived feed of this symposium » The world is changing at a rapid pace. Scientists have documented significant changes during the past century in climate, […] The post Smithsonian Symposium: The Anthropocene appeared first on Smithsonian Insider. Full Article Animals Anthropology Marine Science Research News Science & Nature biodiversity carbon dioxide climate change conservation conservation biology endangered species National Museum of Natural History Smithsonian Conservation Biology Institute
mp Study: Bleaching events impair long-term coral reproduction By insider.si.edu Published On :: Wed, 19 Nov 2014 23:59:00 +0000 A new study by Florida State University and Smithsonian Institution biologists shows that bleaching events brought on by rising sea temperatures are having a detrimental […] The post Study: Bleaching events impair long-term coral reproduction appeared first on Smithsonian Insider. Full Article Animals Marine Science Research News Science & Nature biodiversity carbon dioxide Caribbean climate change conservation conservation biology coral reefs endangered species fishes National Museum of Natural History Tropical Research Institute
mp In modern amphibian “ark,” new species added with due diligence By insider.si.edu Published On :: Mon, 08 Dec 2014 18:07:42 +0000 Bright orange with a distinctive call the Panamanian poison dart frog Andinobates geminisae lives in only a small area of the Caribbean Coast of Panama. […] The post In modern amphibian “ark,” new species added with due diligence appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature amphibian captive breeding Caribbean chytrid fungus climate change conservation biology endangered species frogs Smithsonian's National Zoo
mp Bizarre ocean amphipod has 16 retinas in each eye By insider.si.edu Published On :: Thu, 15 Jan 2015 17:01:37 +0000 Tiny and transparent the marine crustacean Paraphronima gracilis sees the world through two large eyes that envelope its head like a high-tech space helmet. Now, […] The post Bizarre ocean amphipod has 16 retinas in each eye appeared first on Smithsonian Insider. Full Article Animals Marine Science Science & Nature National Museum of Natural History
mp Clouded leopards, from crisis to success: Q&A with Janine Brown By insider.si.edu Published On :: Tue, 31 Mar 2015 14:00:25 +0000 The clouded leopard, a native of Southeast Asia, is among the most charismatic, secretive and least understood cat species in the world. In 2002, the […] The post Clouded leopards, from crisis to success: Q&A with Janine Brown appeared first on Smithsonian Insider. Full Article Animals Q & A Research News Science & Nature biodiversity conservation conservation biology endangered species extinction mammals Smithsonian Conservation Biology Institute Smithsonian's National Zoo veterinary medicine
mp 3 new species of dwarf dragons discovered in Ecuador & Peru By insider.si.edu Published On :: Mon, 06 Apr 2015 13:09:31 +0000 Three new species of dragon-esque woodlizards have been discovered in the Andean cloud forests of Peru and Ecuador. The new species differ from their closest […] The post 3 new species of dwarf dragons discovered in Ecuador & Peru appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature conservation biology National Museum of Natural History new species reptiles South America
mp Complex Organic Molecules Discovered in Infant Star System By insider.si.edu Published On :: Wed, 08 Apr 2015 17:58:34 +0000 For the first time, astronomers have detected the presence of complex organic molecules, the building blocks of life, in a protoplanetary disk surrounding a young […] The post Complex Organic Molecules Discovered in Infant Star System appeared first on Smithsonian Insider. Full Article Research News Science & Nature Space astronomy astrophysics Center for Astrophysics | Harvard & Smithsonian Smithsonian Astrophysical Observatory
mp This simple trick helped fix a wildlife water shortage in Cambodia By insider.si.edu Published On :: Wed, 17 Jun 2015 13:05:17 +0000 It is a simple solution to a wildlife water shortage. During the annual dry season [November through April] in a protected forest in eastern Cambodia, […] The post This simple trick helped fix a wildlife water shortage in Cambodia appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature amphibian biodiversity birds camera traps climate change conservation conservation biology endangered species frogs mammals migratory birds Smithsonian Conservation Biology Institute
mp Warming temperatures may mean more monarch generations in some areas of North America By insider.si.edu Published On :: Wed, 19 Aug 2015 16:25:22 +0000 Warming temperatures may mean more generations of monarch butterflies in North America during summer months, say scientists who recently finished experiments with monarch caterpillars and […] The post Warming temperatures may mean more monarch generations in some areas of North America appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature climate change
mp Tattoos: Telling Stories in the Flesh. Q&A with Lars Krutak By insider.si.edu Published On :: Tue, 25 Aug 2015 16:47:33 +0000 From patterns etched into the skin of an ancient mummy to the colorful designs that adorn people today, tattoos are not just skin deep; they […] The post Tattoos: Telling Stories in the Flesh. Q&A with Lars Krutak appeared first on Smithsonian Insider. Full Article Anthropology History & Culture Science & Nature National Museum of Natural History
mp In Western China’s deserts an ancient competition for water resumes By insider.si.edu Published On :: Tue, 08 Sep 2015 12:11:33 +0000 If you were dumped into the middle of a desert, your first instinct would be to look for water—it is, after all, the stuff of […] The post In Western China’s deserts an ancient competition for water resumes appeared first on Smithsonian Insider. Full Article Animals Research News Science & Nature biodiversity conservation conservation biology endangered species extinction mammals Smithsonian's National Zoo
mp Study reveals ancient link between mammoth dung and pumpkin pie By insider.si.edu Published On :: Mon, 23 Nov 2015 18:16:53 +0000 New research by a team of archaeologists has revealed a curious connection between our traditional Thanksgiving dinner and the taste buds of prehistoric mammoths and […] The post Study reveals ancient link between mammoth dung and pumpkin pie appeared first on Smithsonian Insider. Full Article Anthropology Dinosaurs & Fossils Research News Science & Nature archaeology conservation biology extinction National Museum of Natural History
mp Astronomers complete First Search for Visible Light Associated with Gravitational Waves By insider.si.edu Published On :: Tue, 16 Feb 2016 17:35:22 +0000 Einstein’s general theory of relativity predicts the emission of gravitational waves by massive celestial bodies moving though space-time. For the past century gravitational waves have […] The post Astronomers complete First Search for Visible Light Associated with Gravitational Waves appeared first on Smithsonian Insider. Full Article Research News Science & Nature Space
mp A Purple Accident and Its Vibrant Impact on the Modern World By insider.si.edu Published On :: Wed, 23 Mar 2016 12:07:49 +0000 If you were an average person in the 18th and early 19th centuries, chances are your wardrobe was 50 shades of beige, with maybe a […] The post A Purple Accident and Its Vibrant Impact on the Modern World appeared first on Smithsonian Insider. Full Article Art History & Culture Science & Nature chromatics National Museum of Natural History Smithsonian Libraries
mp Did ripening fruit help hominids develop complex hands? By insider.si.edu Published On :: Thu, 12 May 2016 11:32:13 +0000 One of the primary features that distinguish hominids such as chimpanzees, gorillas and humans from the rest of the animal kingdom are uniquely dexterous hands. […] The post Did ripening fruit help hominids develop complex hands? appeared first on Smithsonian Insider. Full Article Animals Anthropology Research News Science & Nature evolution mammals Smithsonian's National Zoo Tropical Research Institute
mp Billions & Billions of Periodical Cicadas By insider.si.edu Published On :: Fri, 03 Jun 2016 16:45:26 +0000 When soil temperatures 8 inches under the ground reach a balmy 64 degrees Fahrenheit, periodical cicadas emerge on the U.S. East Coast. They tend to […] The post Billions & Billions of Periodical Cicadas appeared first on Smithsonian Insider. Full Article Animals Science & Nature Snapshot insects National Museum of Natural History Smithsonian Libraries
mp Bizarre new marine worms covered in bristles, wrinkles & bumps By insider.si.edu Published On :: Fri, 07 Oct 2016 15:06:56 +0000 An extraordinary arrangement of bristles, wrinkles and wart-like bumps cover the cold skin of Sphaerephesia amphorata, a new deep-sea worm described and named by researchers […] The post Bizarre new marine worms covered in bristles, wrinkles & bumps appeared first on Smithsonian Insider. Full Article Animals Marine Science Science & Nature National Museum of Natural History worms
mp A Planet in Peril: Q&A with Suzan Murray of the Smithsonian Global Health Program By insider.si.edu Published On :: Mon, 07 Nov 2016 20:41:46 +0000 With roughly 5,500 individuals remaining in the wild, the black rhino population is critically endangered. To help save these iconic African giants, at risk for […] The post A Planet in Peril: Q&A with Suzan Murray of the Smithsonian Global Health Program appeared first on Smithsonian Insider. Full Article Animals Science & Nature mammals Smithsonian's National Zoo veterinary medicine
mp By simply pooping, comb jellies expel long-held scientific misconception By insider.si.edu Published On :: Mon, 28 Nov 2016 18:58:16 +0000 In 2012, under a scientist’s watchful eye, a comb jelly ate some fish, digested it, and excreted the waste out its back end. In doing […] The post By simply pooping, comb jellies expel long-held scientific misconception appeared first on Smithsonian Insider. Full Article Animals Marine Science Research News Science & Nature National Museum of Natural History
mp Peacock bass invasion Had devastating, long-term impact on Panama’s Fish By insider.si.edu Published On :: Mon, 12 Dec 2016 18:21:20 +0000 In 1969, 60 to 100 peacock bass imported from Buga, Colombia, were introduced into a pond in Panama for sport fishing. Several individuals escaped. By […] The post Peacock bass invasion Had devastating, long-term impact on Panama’s Fish appeared first on Smithsonian Insider. Full Article Animals Marine Science Research News Science & Nature Spotlight invasive species Smithsonian Tropical Research Institute
mp Paleo-detectives energize great whale mystery: how & when baleen evolved By insider.si.edu Published On :: Fri, 07 Apr 2017 18:20:24 +0000 A bizarre change occurs in the mouth of a humpback whale during its development in the womb. Several dozen tooth buds sprout in a row […] The post Paleo-detectives energize great whale mystery: how & when baleen evolved appeared first on Smithsonian Insider. Full Article Animals Dinosaurs & Fossils Marine Science Research News Science & Nature evolution fossils mammals National Museum of Natural History
mp Surprise: Distinctive new surgeonfish species makes an improbable debut By insider.si.edu Published On :: Wed, 07 Jun 2017 19:10:10 +0000 Sometimes there’s just no telling what will turn up at the local market. Fish biologist Jeff Williams of the Smithsonian’s National Museum of Natural History […] The post Surprise: Distinctive new surgeonfish species makes an improbable debut appeared first on Smithsonian Insider. Full Article Animals Marine Science Q & A Research News Science & Nature biodiversity fishes National Museum of Natural History
mp In search for life, ultraviolet light may be ultra important By insider.si.edu Published On :: Thu, 31 Aug 2017 09:13:51 +0000 In everyday life, ultraviolet, or UV, light earns a bad reputation for being responsible for sunburns and other harmful effects on humans. However, research suggests […] The post In search for life, ultraviolet light may be ultra important appeared first on Smithsonian Insider. Full Article Science & Nature Space Center for Astrophysics | Harvard & Smithsonian Smithsonian Astrophysical Observatory
mp Underpaid women “computers” mapped the universe in the 19th century By insider.si.edu Published On :: Tue, 27 Mar 2018 11:49:04 +0000 Every day, astronomers at the Harvard-Smithsonian Center for Astrophysics depend on computers to help them solve the mysteries of the universe, just as they did […] The post Underpaid women “computers” mapped the universe in the 19th century appeared first on Smithsonian Insider. Full Article Science & Nature Space astrophysics Center for Astrophysics | Harvard & Smithsonian
mp Use GPO to Import Safe Senders & Auto Download External Email Content By www.bleepingcomputer.com Published On :: 2020-04-28T01:51:44-05:00 Full Article
mp Structure of Thermococcus litoralis Δ1-pyrroline-2-carboxylate reductase in complex with NADH and l-proline By scripts.iucr.org Published On :: 2020-04-29 l-Hydroxyproline (l-Hyp) is a nonstandard amino acid that is present in certain proteins, in some antibiotics and in the cell-wall components of plants. l-Hyp is the product of the post-translational modification of protein prolines by prolyl hydroxylase enzymes, and the isomers trans-3-hydroxy-l-proline (T3LHyp) and trans-4-hydroxy-l-proline (T4LHyp) are major components of mammalian collagen. T4LHyp follows two distinct degradation pathways in bacteria and mammals, while T3LHyp is metabolized by a two-step metabolic pathway that is conserved in bacteria and mammals, which involves a T3LHyp dehydratase and a Δ1-pyrroline-2-carboxylate (Pyr2C) reductase. In order to shed light on the structure and catalysis of the enzyme involved in the second step of the T3LHyp degradation pathway, the crystal structure of Pyr2C reductase from the archaeon Thermococcus litoralis DSM 5473 complexed with NADH and l-proline is presented. The model allows the mapping of the residues involved in cofactor and product binding and represents a valid model for rationalizing the catalysis of Pyr2C reductases. Full Article text
mp Structure of the N-terminal domain of ClpC1 in complex with the antituberculosis natural product ecumicin reveals unique binding interactions By scripts.iucr.org Published On :: 2020-04-23 The biological processes related to protein homeostasis in Mycobacterium tuberculosis, the etiologic agent of tuberculosis, have recently been established as critical pathways for therapeutic intervention. Proteins of particular interest are ClpC1 and the ClpC1–ClpP1–ClpP2 proteasome complex. The structure of the potent antituberculosis macrocyclic depsipeptide ecumicin complexed with the N-terminal domain of ClpC1 (ClpC1-NTD) is presented here. Crystals of the ClpC1-NTD–ecumicin complex were monoclinic (unit-cell parameters a = 80.0, b = 130.0, c = 112.0 Å, β = 90.07°; space group P21; 12 complexes per asymmetric unit) and diffracted to 2.5 Å resolution. The structure was solved by molecular replacement using the self-rotation function to resolve space-group ambiguities. The new structure of the ecumicin complex showed a unique 1:2 (target:ligand) stoichiometry exploiting the intramolecular dyad in the α-helical fold of the target N-terminal domain. The structure of the ecumicin complex unveiled extensive interactions in the uniquely extended N-terminus, a critical binding site for the known cyclopeptide complexes. This structure, in comparison with the previously reported rufomycin I complex, revealed unique features that could be relevant for understanding the mechanism of action of these potential antituberculosis drug leads. Comparison of the ecumicin complex and the ClpC1-NTD-L92S/L96P double-mutant structure with the available structures of rufomycin I and cyclomarin A complexes revealed a range of conformational changes available to this small N-terminal helical domain and the minor helical alterations involved in the antibiotic-resistance mechanism. The different modes of binding and structural alterations could be related to distinct modes of action. Full Article text
mp Book Review: Double Exposure: photos of African American History & Culture By insider.si.edu Published On :: Mon, 17 Oct 2016 15:33:08 +0000 Photographs, by virtue of their static nature, not only allow us to look back to a fixed point in time, but also give us a […] The post Book Review: Double Exposure: photos of African American History & Culture appeared first on Smithsonian Insider. Full Article Art Book Review History & Culture Spotlight African American civil rights National Museum of African American History and Culture photography
mp Connecting WiFi dongle to computer running Windows 98 SE By www.bleepingcomputer.com Published On :: 2019-12-07T17:39:29-05:00 Full Article
mp Verifying Mobo components on your prospective buy. By www.bleepingcomputer.com Published On :: 2020-01-08T22:09:08-05:00 Full Article
mp {beta}1 integrin-mediated signaling regulates MT1-MMP phosphorylation to promote tumour cell invasion [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-03-23T03:21:37-07:00 Olivia R. Grafinger, Genya Gorshtein, Tyler Stirling, Megan I. Brasher, and Marc G. CoppolinoMalignant cancer cells can invade extracellular matrix (ECM) through the formation of F-actin-rich subcellular structures termed invadopodia. ECM degradation at invadopodia is mediated by matrix metalloproteinases (MMPs), and recent findings indicate that membrane-anchored membrane type 1-matrix metalloproteinase (MT1-MMP) has a primary role in this process. Maintenance of an invasive phenotype is dependent on internalization of MT1-MMP from the plasma membrane and its recycling to sites of ECM remodeling. Internalization of MT1-MMP is dependent on its phosphorylation, and here we examine the role of β1 integrin-mediated signaling in this process. Activation of β1 integrin using the antibody P4G11 induced phosphorylation and internalization of MT1-MMP and resulted in increased cellular invasiveness and invadopodium formation in vitro. We also observed phosphorylation of Src and epidermal growth factor receptor (EGFR) and an increase in their association in response to β1 integrin activation, and determined that Src and EGFR promote phosphorylation of MT1-MMP on Thr567. These results suggest that MT1-MMP phosphorylation is regulated by a β1 integrin-Src-EGFR signaling pathway that promotes recycling of MT1-MMP to sites of invadopodia formation during cancer cell invasion. Full Article
mp Primary myeloid cell proteomics and transcriptomics: importance of ss tubulin isotypes for osteoclast function [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-04-07T06:16:39-07:00 David Guerit, Pauline Marie, Anne Morel, Justine Maurin, Christel Verollet, Brigitte Raynaud-Messina, Serge Urbach, and Anne BlangyAmong hematopoietic cells, osteoclasts (Oc) and immature dendritic cells (Dc) are closely related myeloid cells with distinct functions; Oc participate skeleton maintenance while Dc sample the environment for foreign antigens. Such specificities rely on profound modifications of gene and protein expression during Oc and Dc differentiation. We provide global proteomic and transcriptomic analyses of primary mouse Oc and Dc, based on original SILAC and RNAseq data. We established specific signatures for Oc and Dc including genes and proteins of unknown functions. In particular, we showed that Oc and Dc have the same α and β tubulin isotypes repertoire but that Oc express much more β tubulin isotype Tubb6. In both mouse and human Oc, we demonstrate that elevated expression of Tubb6 in Oc is necessary for correct podosomes organization and thus for the structure of the sealing zone, which sustains the bone resorption apparatus. Hence, lowering Tubb6 expression hindered Oc resorption activity. Overall, we highlight here potential new regulators of Oc and Dc biology and illustrate the functional importance of the tubulin isotype repertoire in the biology of differentiated cells. Full Article
mp Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes [TOOLS AND RESOURCES] By jcs.biologists.org Published On :: 2020-04-15T01:46:56-07:00 Björn Eismann, Teresa G. Krieger, Jürgen Beneke, Ruben Bulkescher, Lukas Adam, Holger Erfle, Carl Herrmann, Roland Eils, and Christian Conrad3D cell cultures enable the in vitro study of dynamic biological processes such as the cell cycle, but their use in high-throughput screens remains impractical with conventional fluorescent microscopy. Here, we present a screening workflow for the automated evaluation of mitotic phenotypes in 3D cell cultures by light-sheet microscopy. After sample preparation by a liquid handling robot, cell spheroids are imaged for 24 hours in toto with a dual-view inverted selective plane illumination microscope (diSPIM) with a much improved signal-to-noise ratio, higher imaging speed, isotropic resolution and reduced light exposure compared to a spinning disc confocal microscope. A dedicated high-content image processing pipeline implements convolutional neural network based phenotype classification. We illustrate the potential of our approach by siRNA knock-down and epigenetic modification of 28 mitotic target genes for assessing their phenotypic role in mitosis. By rendering light-sheet microscopy operational for high-throughput screening applications, this workflow enables target gene characterization or drug candidate evaluation in tissue-like 3D cell culture models. Full Article
mp Compartmentalization of adenosine metabolism in cancer cells and its modulation during acute hypoxia [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-04-21T05:32:09-07:00 Karolina Losenkova, Mariachiara Zuccarini, Marika Karikoski, Juha Laurila, Detlev Boison, Sirpa Jalkanen, and Gennady G. YegutkinExtracellular adenosine mediates diverse anti-inflammatory, angiogenic and vasoactive effects and becomes an important therapeutic target for cancer, which has been translated into clinical trials. This study was designed to comprehensively assess adenosine metabolism in prostate and breast cancer cells. We identified cellular adenosine turnover as a complex cascade, comprised of (a) the ectoenzymatic breakdown of ATP via sequential nucleotide pyrophosphatase/phosphodiesterase-1, ecto-5’-nucleotidase/CD73 and adenosine deaminase reactions, and ATP re-synthesis through counteracting adenylate kinase and nucleoside diphosphokinase; (b) the uptake of nucleotide-derived adenosine via equilibrative nucleoside transporters; and (c) the intracellular adenosine phosphorylation into ATP by adenosine kinase and other nucleotide kinases. The exposure of cancer cells to 1% O2 for 24 hours triggered ~2-fold up-regulation of CD73, without affecting nucleoside transporters, adenosine kinase activity and cellular ATP content. The ability of adenosine to inhibit the tumor-initiating potential of breast cancer cells via receptor-independent mechanism was confirmed in vivo using a xenograft mouse model. The existence of redundant pathways controlling extracellular and intracellular adenosine provides a sufficient justification for reexamination of the current concepts of cellular purine homeostasis and signaling in cancer. Full Article
mp A stable core of GCPs 4, 5 and 6 promotes the assembly of {gamma}-tubulin ring complexes [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-04-21T05:32:09-07:00 Laurence Haren, Dorian Farache, Laurent Emorine, and Andreas Merdes-tubulin is a major protein involved in the nucleation of microtubules in all eukaryotes. It forms two different complexes with proteins of the GCP family (gamma-tubulin complex proteins): -tubulin small complexes (TuSCs), containing -tubulin and GCPs 2 and 3, and -tubulin ring complexes (TuRCs), containing multiple TuSCs, in addition to GCPs 4, 5, and 6. Whereas the structure and assembly properties of TuSCs have been intensively studied, little is known about the assembly of TuRCs, and about the specific roles of GCPs 4, 5, and 6. Here, we demonstrate that two copies of GCP4 and one copy each of GCP5 and GCP6 form a salt-resistant sub-complex within the TuRC that assembles independently of the presence of TuSCs. Incubation of this sub-complex with cytoplasmic extracts containing TuSCs leads to the reconstitution of TuRCs that are competent to nucleate microtubules. In addition, we investigate sequence extensions and insertions that are specifically found at the amino-terminus of GCP6, and between the GCP6 grip1 and grip2 motifs, and we demonstrate that these are involved in the assembly or stabilization of the TuRC. Full Article
mp Tetrahymena Poc5 is a transient basal body component that is important for basal body maturation [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-04-29T02:35:34-07:00 Westley Heydeck, Brian A. Bayless, Alexander J. Stemm-Wolf, Eileen T. O'Toole, Amy S. Fabritius, Courtney Ozzello, Marina Nguyen, and Mark WineyBasal bodies (BBs) are microtubule-based organelles that template and stabilize cilia at the cell surface. Centrins ubiquitously associate with BBs and function in BB assembly, maturation, and stability. Human POC5 (hPOC5) is a highly conserved centrin-binding protein that binds centrins through Sfi1p-like repeats and is required for building full-length, mature centrioles. Here, we use the BB-rich cytoskeleton of Tetrahymena thermophila to characterize Poc5 BB functions. Tetrahymena Poc5 (TtPoc5) uniquely incorporates into assembling BBs and is then removed from mature BBs prior to ciliogenesis. Complete genomic knockout of TtPOC5 leads to a significantly increased production of BBs yet a markedly reduced ciliary density, both of which are rescued by reintroduction of TtPoc5. A second Tetrahymena POC5-like gene, SFR1, is similarly implicated in modulating BB production. When TtPOC5 and SFR1 are co-deleted, cell viability is compromised, and levels of BB overproduction are exacerbated. Overproduced BBs display defective transition zone formation and a diminished capacity for ciliogenesis. This study uncovers a requirement for Poc5 in building mature BBs, providing a possible functional link between hPOC5 mutations and impaired cilia. Full Article
mp BMP4 promotes the metastasis of gastric cancer by inducing epithelial-mesenchymal transition via Id1 [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-05-06T08:30:40-07:00 Ganlu Deng, Yihong Chen, Cao Guo, Ling Yin, Ying Han, Yiyi Li, Yaojie Fu, Changjing Cai, Hong Shen, and Shan ZengEpithelial-mesenchymal transition (EMT) is a crucial process for cancer cells to acquire metastatic potential, which primarily causes death in gastric cancer (GC) patients. Bone morphogenetic protein 4 (BMP4) is a member of the TGF-β family that plays an indispensable role in human cancers. However, little is known about its roles in GC metastasis. In this study, BMP4 was found to be frequently overexpressed in GC tissues and was correlated with patient's poor prognosis. BMP4 was upregulated in GC cell lines and promoted EMT and metastasis of GC cells both in vitro and in vivo, while knockdown of BMP4 significantly inhibited EMT and metastasis of GC cells. Meanwhile, the inhibitor of DNA binding 1 (Id1) was identified as a downstream target of BMP4 by PCR arrays and upregulated via Smad1/5/8 phosphorylation. Id1 knockdown attenuated BMP4-induced EMT and invasion in GC cells. Moreover, Id1 overexpression in BMP4 knockdown cells restored the promotion of EMT and cell invasion. In summary, BMP4 induced EMT to promote GC metastasis by upregulating Id1 expression. Antagonizing BMP4 may be a potential therapeutic strategy in GC metastasis. Full Article
mp OPTN recruitment to a Golgi-proximal compartment regulates immune signalling and cytokine secretion [RESEARCH ARTICLE] By jcs.biologists.org Published On :: 2020-05-06T08:30:40-07:00 Thomas O'Loughlin, Antonina J. Kruppa, Andre L. R. Ribeiro, James R. Edgar, Abdulaziz Ghannam, Andrew M. Smith, and Folma BussOptineurin (OPTN) is a multifunctional protein involved in autophagy, secretion as well as NF-B and IRF3 signalling and OPTN mutations are associated with several human diseases. Here we show that, in response to viral RNA, OPTN translocates to foci in the perinuclear region, where it negatively regulates NF-B and IRF3 signalling pathways and downstream pro-inflammatory cytokine secretion. These OPTN foci consist of a tight cluster of small membrane vesicles, which are positive for ATG9A. Disease mutations linked to POAG cause aberrant foci formation in the absence of stimuli, which correlates with the ability of OPTN to inhibit signalling. Using proximity labelling proteomics, we identify the LUBAC complex, CYLD and TBK1 as part of the OPTN interactome and show that these proteins are recruited to this OPTN-positive perinuclear compartment. Our work uncovers a crucial role for OPTN in dampening NF-B and IRF3 signalling through the sequestration of LUBAC and other positive regulators in this viral RNA-induced compartment leading to altered pro-inflammatory cytokine secretion. Full Article