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Reconciling the regulatory role of Munc18 proteins in SNARE-complex assembly

Mammalian Munc18 proteins are essential for membrane fusion and human health. Here, we review the literature describing structural and in vitro data, and identify a possible explanation for the conflicting functional roles that have been reported.






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Solution structure and assembly of β-amylase 2 from Arabidopsis thaliana

Solution structure of β-amylase 2 from Arabidopsis thaliana shows the role of the conserved N-terminus in enzyme tetramer formation.




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Six 1-aroyl-4-(4-meth­oxy­phen­yl)piperazines: similar mol­ecular structures but different patterns of supra­molecular assembly

Six new 1-aroyl-4-(4-meth­oxy­phen­yl)piperazines have been prepared, using coupling reactions between benzoic acids and N-(4-meth­oxy­phen­yl)piperazine. There are no significant hydrogen bonds in the structure of 1-benzoyl-4-(4-meth­oxy­phen­yl)piperazine, C18H20N2O2, (I). The mol­ecules of 1-(2-fluoro­benzo­yl)-4-(4-meth­oxy­phen­yl)piperazine, C18H19FN2O2, (II), are linked by two C—H⋯O hydrogen bonds to form chains of rings, which are linked into sheets by an aromatic π–π stacking inter­action. 1-(2-Chloro­benzo­yl)-4-(4-meth­oxy­phen­yl)piperazine, C18H19ClN2O2, (III), 1-(2-bromo­benzo­yl)-4-(4-meth­oxy­phen­yl)piperazine, C18H19BrN2O2, (IV), and 1-(2-iodo­benzo­yl)-4-(4-meth­oxyphen­yl)piperazine, C18H19IN2O2, (V), are isomorphous, but in (III) the aroyl ring is disordered over two sets of atomic sites having occupancies of 0.942 (2) and 0.058 (2). In each of (III)–(V), a combination of two C—H⋯π(arene) hydrogen bonds links the mol­ecules into sheets. A single O—H⋯O hydrogen bond links the mol­ecules of 1-(2-hy­droxy­benzo­yl)-4-(4-meth­oxy­phen­yl)piperazine, C18H20N2O3, (VI), into simple chains. Comparisons are made with the structures of some related compounds.




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Twelve 4-(4-meth­oxy­phen­yl)piperazin-1-ium salts containing organic anions: supra­molecular assembly in one, two and three dimensions

Twelve 4-(4-meth­oxy­phen­yl)piperazin-1-ium salts containing organic anions have been prepared and structurally characterized. The monohydrated benzoate, 4-fluoro­benzoate, 4-chloro­benzoate and 4-bromo­benzoate salts, C11H17N2O+·C7H5O2−·H2O (I), C11H17N2O+·C7H4FO2−·H2O (II), C11H17N2O+·C7H4ClO2−·H2O (III), and C11H17N2O+·C7H4BrO2−·H2O (IV), respectively, are isomorphous and all exhibit disorder in the 4-meth­oxy­phenyl unit: the components are linked by N—H⋯O and O—H⋯O hydrogen bond to form chains of rings. The unsolvated 2-hy­droxy­benzoate, pyridine-3-carboxyl­ate and 2-hy­droxy-3,5-di­nitro­benzoate salts, C11H17N2O+·C7H5O3− (V), C11H17N2O+·C6H4NO2− (VI) and C11H17N2O+·C7H3N2O7− (VII), respectively, are all fully ordered: the components of (V) are linked by multiple N—H⋯O hydrogen bonds to form a chain of rings; those of (VI) are linked into a three-dimensional framework by a combination of N—H⋯O, C—H⋯O and C—H⋯N hydrogen bonds and those of (VII), where the anion has a structure reminiscent of the picrate anion, are linked into a three-dimensional array by N—H⋯O and C—H⋯O hydrogen bonds. The hydrogensuccinate and hydrogenfumarate salts, C11H17N2O+·C4H5O4− (VIII) and C11H17N2O+·C4H3O3− (IX), respectively, are isomorphous, and both exhibit disorder in the anionic component: N—H⋯O and O—H⋯O hydrogen bonds link the ions into sheets, which are further linked by C—H⋯π(arene) inter­actions. The anion of the hydrogenmaleate salt, C11H17N2O+·C4H3O3− (X), contains a very short and nearly symmetrical O⋯H⋯O hydrogen bond, and N—H⋯O hydrogen bonds link the anions into chains of rings. The ions in the tri­chloro­acetate salt, C11H17N2O+·C2Cl3O2− (XI), are linked into simple chains by N—H⋯O hydrogen bonds. In the hydrated chloranilate salt, 2C11H17N2O+·C6Cl2O42−·2H2O (XII), which crystallizes as a non-merohedral twin, the anion lies across a centre of inversion in space group P21/n, and a combination of N—H⋯O and O—H⋯O hydrogen bonds generates complex sheets. Comparisons are made with the structures of some related compounds.




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Two isostructural 3-(5-ar­yloxy-3-methyl-1-phenyl-1H-pyrazol-4-yl)-1-(thio­phen-2-yl)prop-2-en-1-ones: disorder and supra­molecular assembly

Two new chalcones containing both pyrazole and thio­phene substituents have been prepared and structurally characterized. 3-(3-Methyl-5-phen­oxy-1-phenyl-1H-pyrazol-4-yl)-1-(thio­phen-2-yl)prop-2-en-1-one, C23H18N2O2S (I), and 3-[3-methyl-5-(2-methyl­phen­oxy)-1-phenyl-1H-pyrazol-4-yl]-1-(thio­phen-2-yl)prop-2-en-1-one, C24H20N2O2S (II), are isomorphous as well as isostructural, and in each the thio­phene substituent is disordered over two sets of atomic sites having occupancies 0.844 (3) and 0.156 (3) in (I), and 0.883 (2) and 0.117 (2) in (II). In each structure, the mol­ecules are linked into sheets by a combination of C—H⋯N and C—H⋯O hydrogen bonds. Comparisons are made with some related compounds.




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Crystal structure of a two-dimensional metal–organic framework assembled from lithium(I) and γ-cyclo­dextrin

The crystal structure of the polymeric title compound, catena-poly[[[di­aqua­lithium]-μ-γ-cyclo­dextrin(1−)-[aqua­lithium]-μ-γ-cyclo­dextrin(1−)] pentadecahydrate], {[Li2(C48H79O40)2(H2O)3]·15H2O}n, consists of deprotonated γ-cyclo­dextrin (CD) mol­ecules assembled by lithium ions into metal–organic ribbons that are cross-linked by multiple O—H⋯O hydrogen bonds into sheets extending parallel to (0overline11). Within a ribbon, one Li+ ion is coordinated by one deprotonated hydroxyl group of the first γ-CD torus and by one hydroxyl group of the second γ-CD torus as well as by two water mol­ecules. The other Li+ ion is coordinated by one deprotonated hydroxyl and by one hydroxyl group of the second γ-CD torus, by one hydroxyl group of the first γ-CD torus as well as by one water mol­ecule. The coordination spheres of both Li+ cations are distorted tetra­hedral. The packing of the structure constitute channels along the a axis. Parts of the hy­droxy­methyl groups in cyclo­dextrin molecules as well as water mol­ecules show two-component disorder. Electron density associated with additional disordered solvent mol­ecules inside the cavities was removed with the SQUEEZE [Spek (2015). Acta Cryst. C71, 9–18] routine in PLATON. These solvent mol­ecules are not considered in the given chemical formula and other crystal data. Five out of the sixteen hy­droxy­methyl groups and one water mol­ecule are disordered over two sets of sites.




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Structural insights into stressosome assembly

The stressosome transduces environmental stress signals to SigB to upregulate SigB-dependent transcription, which is required for bacterial viability. The stressosome core is composed of RsbS and at least one of the RsbR paralogs. A previous cryo-electron microscopy (cryo-EM) structure of the RsbRA–RsbS complex determined under a D2 symmetry restraint showed that the stressosome core forms a pseudo-icosahedron consisting of 60 STAS domains of RsbRA and RsbS. However, it is still unclear how RsbS and one of the RsbR paralogs assemble into the stressosome. Here, an assembly model of the stressosome is presented based on the crystal structure of the RsbS icosahedron and cryo-EM structures of the RsbRA–RsbS complex determined under diverse symmetry restraints (nonsymmetric C1, dihedral D2 and icosahedral I envelopes). 60 monomers of the crystal structure of RsbS fitted well into the I-restrained cryo-EM structure determined at 4.1 Å resolution, even though the STAS domains in the I envelope were averaged. This indicates that RsbS and RsbRA share a highly conserved STAS fold. 22 protrusions observed in the C1 envelope, corresponding to dimers of the RsbRA N-domain, allowed the STAS domains of RsbRA and RsbS to be distinguished in the stressosome core. Based on these, the model of the stressosome core was reconstructed. The mutation of RsbRA residues at the binding interface in the model (R189A/Q191A) significantly reduced the interaction between RsbRA and RsbS. These results suggest that nonconserved residues in the conserved STAS folds between RsbS and RsbR paralogs determine stressosome assembly.




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Consistency and variability of cocrystals containing positional isomers: the self-assembly evolution mechanism of supramolecular synthons of cresol–piperazine

The disposition of functional groups can induce variations in the nature and type of interactions and hence affect the molecular recognition and self-assembly mechanism in cocrystals. To better understand the formation of cocrystals on a molecular level, the effects of disposition of functional groups on the formation of cocrystals were systematically and comprehensively investigated using cresol isomers (o-, m-, p-cresol) as model compounds. Consistency and variability in these cocrystals containing positional isomers were found and analyzed. The structures, molecular recognition and self-assembly mechanism of supramolecular synthons in solution and in their corresponding cocrystals were verified by a combined experimental and theoretical calculation approach. It was found that the heterosynthons (heterotrimer or heterodimer) combined with O—H⋯N hydrogen bonding played a significant role. Hirshfeld surface analysis and computed interaction energy values were used to determine the hierarchical ordering of the weak interactions. The quantitative analyses of charge transfers and molecular electrostatic potential were also applied to reveal and verify the reasons for consistency and variability. Finally, the molecular recognition, self-assembly and evolution process of the supramolecular synthons in solution were investigated. The results confirm that the supramolecular synthon structures formed initially in solution would be carried over to the final cocrystals, and the supramolecular synthon structures are the precursors of cocrystals and the information memory of the cocrystallization process, which is evidence for classical nucleation theory.




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The active form of quinol-dependent nitric oxide reductase from Neisseria meningitidis is a dimer

Neisseria meningitidis is carried by nearly a billion humans, causing developmental impairment and over 100 000 deaths a year. A quinol-dependent nitric oxide reductase (qNOR) plays a critical role in the survival of the bacterium in the human host. X-ray crystallographic analyses of qNOR, including that from N. meningitidis (NmqNOR) reported here at 3.15 Å resolution, show monomeric assemblies, despite the more active dimeric sample being used for crystallization. Cryo-electron microscopic analysis of the same chromatographic fraction of NmqNOR, however, revealed a dimeric assembly at 3.06 Å resolution. It is shown that zinc (which is used in crystallization) binding near the dimer-stabilizing TMII region contributes to the disruption of the dimer. A similar destabilization is observed in the monomeric (∼85 kDa) cryo-EM structure of a mutant (Glu494Ala) qNOR from the opportunistic pathogen Alcaligenes (Achromobacter) xylosoxidans, which primarily migrates as a monomer. The monomer–dimer transition of qNORs seen in the cryo-EM and crystallographic structures has wider implications for structural studies of multimeric membrane proteins. X-ray crystallographic and cryo-EM structural analyses have been performed on the same chromatographic fraction of NmqNOR to high resolution. This represents one of the first examples in which the two approaches have been used to reveal a monomeric assembly in crystallo and a dimeric assembly in vitrified cryo-EM grids. A number of factors have been identified that may trigger the destabilization of helices that are necessary to preserve the integrity of the dimer. These include zinc binding near the entry of the putative proton-transfer channel and the preservation of the conformational integrity of the active site. The mutation near the active site results in disruption of the active site, causing an additional destabilization of helices (TMIX and TMX) that flank the proton-transfer channel helices, creating an inert monomeric enzyme.




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Crystal structures of the Bacillus subtilis prophage lytic cassette proteins XepA and YomS

As part of the Virus-X Consortium that aims to identify and characterize novel proteins and enzymes from bacteriophages and archaeal viruses, the genes of the putative lytic proteins XepA from Bacillus subtilis prophage PBSX and YomS from prophage SPβ were cloned and the proteins were subsequently produced and functionally characterized. In order to elucidate the role and the molecular mechanism of XepA and YomS, the crystal structures of these proteins were solved at resolutions of 1.9 and 1.3 Å, respectively. XepA consists of two antiparallel β-sandwich domains connected by a 30-amino-acid linker region. A pentamer of this protein adopts a unique dumbbell-shaped architecture consisting of two discs and a central tunnel. YomS (12.9 kDa per monomer), which is less than half the size of XepA (30.3 kDa), shows homology to the C-terminal part of XepA and exhibits a similar pentameric disc arrangement. Each β-sandwich entity resembles the fold of typical cytoplasmic membrane-binding C2 domains. Only XepA exhibits distinct cytotoxic activity in vivo, suggesting that the N-terminal pentameric domain is essential for this biological activity. The biological and structural data presented here suggest that XepA disrupts the proton motive force of the cytoplasmatic membrane, thus supporting cell lysis.




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Insight into the role of pre-assembly and desolvation in crystal nucleation: a case of p-nitro­benzoic acid

As one of the most important phenomena in crystallization, the crystal nucleation process has always been the focus of research. In this work, influences of pre-assembly species and the desolvation process on the crystal nucleation process were studied. p-Nitro­benzoic acid (PNBA) was taken as a model compound to investigate the relationship between solution chemistry and nucleation kinetics in seven different solvents. One unsolvated form and four solvates of PNBA were obtained and one of the solvates was newly discovered. The nucleation behaviours and nucleation kinetics of PNBA in the seven solvents were studied and analyzed. Density functional theory (DFT) and solvation energy calculation were adopted to evaluate the strength of solute–solvent interactions. Vibrational spectroscopy combined with molecular simulation was applied to reveal the pre-assembly species in the solution. Based on these results, a comprehensive understanding of the relationship between molecular structure, crystal structure, solution chemistry and nucleation dynamics was proposed and discussed. It was found that the structural similarity between solution chemistry and crystal structure, the interaction between specific sites and the overall strength of solvation will jointly affect the nucleation process.




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PyMDA: microcrystal data assembly using Python

The recent developments at microdiffraction X-ray beamlines are making microcrystals of macromolecules appealing subjects for routine structural analysis. Microcrystal diffraction data collected at synchrotron microdiffraction beamlines may be radiation damaged with incomplete data per microcrystal and with unit-cell variations. A multi-stage data assembly method has previously been designed for microcrystal synchrotron crystallography. Here the strategy has been implemented as a Python program for microcrystal data assembly (PyMDA). PyMDA optimizes microcrystal data quality including weak anomalous signals through iterative crystal and frame rejections. Beyond microcrystals, PyMDA may be applicable for assembling data sets from larger crystals for improved data quality.




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Local orientational order in self-assembled nanoparticle films: the role of ligand composition and salt

An X-ray cross-correlation study of the impact of ligand composition and salt content on the self-assembly of soft-shell nanoparticles is presented, indicating symmetry-selective formation of order.




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The Philosophy of Science – A Companion. Edited by Anouk Baberousse, Denis Bonnay and Mikael Cozic. Oxford University Press, 2018. Pp. 768. Price GBP 64.00. ISBN-13 9780190690649.

Book review




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sasPDF: pair distribution function analysis of nanoparticle assemblies from small-angle scattering data

The sasPDF method, an extension of the atomic pair distribution function (PDF) analysis to the small-angle scattering (SAS) regime, is presented. The method is applied to characterize the structure of nanoparticle assemblies with different levels of structural order.




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Clay vessels by Native American potter Jeri Redcorn added to Smithsonian collections

The Caddo people of Arkansas, Louisiana, Texas and Oklahoma have maintained many of their traditional ways and actively work to preserve their unique tribal cultural today. One example is the pottery of Jeri Redcorn.

The post Clay vessels by Native American potter Jeri Redcorn added to Smithsonian collections appeared first on Smithsonian Insider.




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Scientists discover the largest assembly of whale sharks ever recorded

This new research, which involved both surface and aerial surveys, has revealed an enormous aggregation of whale sharks—the largest ever reported—with up to 420 individuals off the coast of the Yucatán Peninsula in Mexico.

The post Scientists discover the largest assembly of whale sharks ever recorded appeared first on Smithsonian Insider.




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X-Class flares released by the Sun, March 6, captured by Atmospheric Imaging Assembly

The Sun’s Active Region 1429 has been shooting off flares and coronal mass ejections since it rotated into Earth’s view on March 2, 2012. Two X-class flares have been released overnight, an X1.3 and an X5.4.

The post X-Class flares released by the Sun, March 6, captured by Atmospheric Imaging Assembly appeared first on Smithsonian Insider.






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Forest corridors essential to gene flow in India’s leopard and tiger populations

As economic expansion and development fragments the forest landscape of central India, the species that rely on that habitat—including endangered tigers and leopards—face dwindling populations […]

The post Forest corridors essential to gene flow in India’s leopard and tiger populations appeared first on Smithsonian Insider.





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“The State of the Birds” assesses health of nation’s birds

One hundred years after the extinction of the passenger pigeon, the nation’s top bird science and conservation groups have come together to publish The State […]

The post “The State of the Birds” assesses health of nation’s birds appeared first on Smithsonian Insider.




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In Belize, Critically endangered wrasse now favorite food of invasive lionfish

Scientists examining the stomach contents of invasive lionfish caught on the inner barrier reef of Belize have discovered that nearly half of the diet of […]

The post In Belize, Critically endangered wrasse now favorite food of invasive lionfish appeared first on Smithsonian Insider.




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In the wild, biodiversity’s power surpasses what experiments predict

Hundreds of experiments have shown biodiversity fosters healthier, more productive ecosystems. But many experts doubted whether these experiments would hold up in the real world. […]

The post In the wild, biodiversity’s power surpasses what experiments predict appeared first on Smithsonian Insider.




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The real history behind science fiction’s ‘2001: A Space Odyssey’

When “2001: A Space Odyssey” premiered April 2, 1968 at Washington, D.C.’s Uptown Theater—not far from the Smithsonian’s National Air and Space Museum—not everyone was […]

The post The real history behind science fiction’s ‘2001: A Space Odyssey’ appeared first on Smithsonian Insider.



  • Art
  • History & Culture
  • Science & Nature
  • Space
  • National Air and Space Museum

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SNAP29 mediates the assembly of histidine-induced CTP synthase filaments in proximity to the cytokeratin network [RESEARCH ARTICLE]

Archan Chakraborty, Wei-Cheng Lin, Yu-Tsun Lin, Kuang-Jing Huang, Pei-Yu Wang, Yi-Feng Chang, Hsiang-Iu Wang, Kung-Ting Ma, Chun-Yen Wang, Xuan-Rong Huang, Yen-Hsien Lee, Bi-Chang Chen, Ya-Ju Hsieh, Kun-Yi Chien, Tzu-Yang Lin, Ji-Long Liu, Li-Ying Sung, Jau-Song Yu, Yu-Sun Chang, and Li-Mei Pai

Under metabolic stress, cellular components can assemble into distinct membraneless organelles for adaptation. One such example is cytidine 5'-triphosphate synthase (CTPS), which forms filamentous structures under glutamine deprivation. We have previously demonstrated that histidine (His)-mediated methylation regulates the formation of CTPS filaments to suppress enzymatic activity and preserve the CTPS protein under Gln deprivation, which promotes cancer cell growth after stress alleviation. However, it remains unclear where and how these enigmatic structures are assembled. Using CTPS-APEX2-mediated in vivo proximity labeling, we found that SNAP29 regulates the spatiotemporal filament assembly of CTPS along the cytokeratin network in a keratin 8 (KRT8)-dependent manner. Knockdown of synaptosome-associated protein 29 (SNAP29) interfered with assembly and relaxed the filament-induced suppression of CTPS enzymatic activity. Furthermore, APEX2 proximity labeling of keratin 18 (KRT18) revealed a spatiotemporal association of SNAP29 with cytokeratin in response to stress. Super-resolution imaging suggests that during CTPS filament formation, SNAP29 interacts with CTPS along the cytokeratin network. This study links the cytokeratin network to the regulation of metabolism by compartmentalization of metabolic enzymes during nutrient deprivation.




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Canonical nucleators are dispensable for stress granule assembly in intestinal progenitors [RESEARCH ARTICLE]

Kasun Buddika, Ishara S. Ariyapala, Mary A. Hazuga, Derek Riffert, and Nicholas S. Sokol

Stressed cells downregulate translation initiation and assemble membrane-less foci termed stress granules (SGs). Extensively characterized in cultured cells, the existence of such structures in stressed adult stem cell pools remain poorly characterized. Here we report that Drosophila orthologs of mammalian SG components AGO1, ATX2, CAPRIN, eIF4E, FMRP, G3BP, LIN-28, PABP, and TIAR are enriched in adult intestinal progenitor cells where they accumulate in small cytoplasmic messenger ribonucleoprotein complexes (mRNPs). Treatment with sodium arsenite or rapamycin reorganized these mRNPs into large cytoplasmic granules. Formation of these intestinal progenitor stress granules (IPSGs) depended on polysome disassembly, led to translational downregulation, and was reversible. While canonical SG nucleators ATX2 and G3BP were sufficient for IPSG formation in the absence of stress, neither of them, nor TIAR, either individually or collectively, were required for stress-induced IPSG formation. This work therefore finds that IPSGs do not assemble via a canonical mechanism, raising the possibility that other stem cell populations employ a similar stress-response mechanism.




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Control of assembly of extra-axonemal structures: the paraflagellar rod of trypanosomes [RESEARCH ARTICLE]

Aline A. Alves, Heloisa B. Gabriel, Maria J. R. Bezerra, Wanderley de Souza, Sue Vaughan, Narcisa L. Cunha-e-Silva, and Jack D. Sunter

Eukaryotic flagella are complex microtubule based organelles and in many organisms there are extra-axonemal structures present, including the outer dense fibres of mammalian sperm and the paraflagellar rod (PFR) of trypanosomes. Flagellum assembly is a complex process occurring across three main compartments, the cytoplasm, the transition fibre-transition zone, and the flagellum. It begins with translation of protein components, followed by their sorting and trafficking into the flagellum, transport to the assembly site and then incorporation. Flagella are formed from over 500 proteins; the principles governing axonemal component assembly are relatively clear. However, the coordination and sites of extra-axonemal structure assembly processes are less clear.

We have discovered two cytoplasmic proteins in T. brucei that are required for PFR formation, PFR assembly factors 1 and 2. Deletion of either PFR-AF1 or PFR-AF2 dramatically disrupted PFR formation and caused a reduction in the amount of major PFR proteins. The presence of cytoplasmic factors required for PFR formation aligns with the concept of processes occurring across multiple compartments to facilitate axoneme assembly and this is likely a common theme for extra-axonemal structure assembly.




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A stable core of GCPs 4, 5 and 6 promotes the assembly of {gamma}-tubulin ring complexes [RESEARCH ARTICLE]

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.




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Kinesin-14s and microtubule dynamics define fission yeast mitotic and meiotic spindle assembly and elongation [RESEARCH ARTICLE]

Ana Loncar, Sergio A. Rincon, Manuel Lera Ramirez, Anne Paoletti, and Phong T. Tran

To segregate the chromosomes faithfully during cell division, cells assemble a spindle that captures the kinetochores and pulls them towards opposite poles. Proper spindle function requires correct interplay between microtubule motors and non-motor proteins. Defects in spindle assembly or changes in spindle dynamics are associated with diseases like cancer or developmental disorders. Here we compared mitotic and meiotic spindles in fission yeast. We show that even though mitotic and meiotic spindles undergo the typical three phases of spindle elongation, they have distinct features. We found that the relative concentration of kinesin-14 Pkl1 is decreased in meiosis I compared to mitosis, while the concentration of kinesin-5 Cut7 remains constant. We identified the second kinesin-14 Klp2 and microtubule dynamics as factors necessary for proper meiotic spindle assembly. This work defines differences between mitotic and meiotic spindles in fission yeast, and provides prospect for future comparative studies.




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Cdc24 interacts with the septins to create a positive feedback during bud site assembly in yeast [RESEARCH ARTICLE]

Julian Chollet, Alexander Dünkler, Anne Bäuerle, Laura Vivero-Pol, Medhanie A. Mulaw, Thomas Gronemeyer, and Nils Johnsson

Yeast cells select the position of their new bud at the beginning of each cell cycle. The recruitment of the septins to this prospective bud site is one of the critical events in a complex assembly pathway that culminates in the outgrowth of a new daughter cell. Hereby, the septin-rods follow the high concentration of Cdc42GTP that is generated by the focused localization of its GEF Cdc24. We show that shortly before budding Cdc24 not only activates Cdc42 but also transiently interacts with Cdc11, the septin subunit that caps both ends of the septin rods. Mutations in Cdc24 reducing the affinity to Cdc11 impair septin recruitment and decrease the stability of the polarity patch. The interaction between septins and Cdc24 thus reinforces bud assembly at sites where septin structures are formed. Once the septins polymerize into the ring, Cdc24 is found at the cortex of the bud and directs its further outgrowth from this position.




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Isotopy classes for 3-periodic net embeddings

Entangled embedded periodic nets and crystal frameworks are defined, along with their dimension type, homogeneity type, adjacency depth and periodic isotopy type. Periodic isotopy classifications are obtained for various families of embedded nets with small quotient graphs. The 25 periodic isotopy classes of depth-1 embedded nets with a single-vertex quotient graph are enumerated. Additionally, a classification is given of embeddings of n-fold copies of pcu with all connected components in a parallel orientation and n vertices in a repeat unit, as well as demonstrations of their maximal symmetry periodic isotopes. The methodology of linear graph knots on the flat 3-torus [0,1)3 is introduced. These graph knots, with linear edges, are spatial embeddings of the labelled quotient graphs of an embedded net which are associated with its periodicity bases.




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How To Email To Multiple Addresses At The Same Time




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Scientists at a new National Zoo laboratory use DNA to diagnose illnesses and discover new species of animals

Researchers at the Smithsonian's National Zoo's new genetics lab use animal DNA to diagnose new diseases, discover new species help in conservation efforts and solve mysteries.

The post Scientists at a new National Zoo laboratory use DNA to diagnose illnesses and discover new species of animals appeared first on Smithsonian Insider.





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Great Cats curator Craig Saffoe discusses his work caring for the National Zoo’s seven frisky lion cubs

What does it take to care for a pride of lions? Go behind the scenes with Great Cats Curator Craig Saffoe as he works with animal keepers and veterinarians to prepare the National Zoo's frisky lion cubs for their public debut. For more about the Zoo's growing pride and to watch them on live webcams: http://nationalzoo.si.edu/Animals/GreatCats/default.cfm?cam=LC4

The post Great Cats curator Craig Saffoe discusses his work caring for the National Zoo’s seven frisky lion cubs appeared first on Smithsonian Insider.




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Roland Kays, curator of mammals at the New York State Museum, discusses a new Smithsonian Website for sharing camera-trap images of wild animals

The post Roland Kays, curator of mammals at the New York State Museum, discusses a new Smithsonian Website for sharing camera-trap images of wild animals appeared first on Smithsonian Insider.





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Spectacular June 7 solar flare seen through the Atmospheric Imaging Assembly aboard Solar Dynamics Observatory

On June 7 the Sun unleashed an spectacular solar flare with a substantial coronal mass ejection. A large cloud of plasma mushroomed up, and while some parts fell back into the Sun, most rushed off into space. The first two segments of this video are seen through the Atmospheric Imaging Assembly aboard NASA's Solar Dynamics Observatory. The AIA was developed by Smithsonian scientists.

The post Spectacular June 7 solar flare seen through the Atmospheric Imaging Assembly aboard Solar Dynamics Observatory appeared first on Smithsonian Insider.




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Tom Crouch, Senior curator in the National Air and Space Museum’s Aeronautics Division, discusses Thaddeus Lowe and the birth of American aerial reconnaissance

Tom Crouch, Senior curator in the National Air and Space Museum's Aeronautics Division, discusses Thaddeus Lowe and the birth of American aerial reconnaissance during the Civil War. This presentation was recorded on May 11, 2011 on the National Mall.

The post Tom Crouch, Senior curator in the National Air and Space Museum’s Aeronautics Division, discusses Thaddeus Lowe and the birth of American aerial reconnaissance appeared first on Smithsonian Insider.




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Kari Bruwelheide, forensic anthropologist at the Smithsonian’s National Museum of Natural History, discusses the power of bones.

Kari Bruwelheide, forensic anthropologist at the Smithsonian's National Museum of Natural History, discusses how she came to work at the museum, the power of human remains and the information that bones can contain. She and her colleagues continue to discover new ways to interpret evidence from bones and burials.

The post Kari Bruwelheide, forensic anthropologist at the Smithsonian’s National Museum of Natural History, discusses the power of bones. appeared first on Smithsonian Insider.



  • Video
  • National Museum of Natural History



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One way businesses are avoiding health care coverage for employees

Business Update with Mark Lacter

Businesses are cutting back on hours to avoid having to provide health care coverage under the new Affordable Care Act.

Steve Julian: Business analyst Mark Lacter, who's affected here?

Mark Lacter: Thirty hours a week is the magic number for workers to be considered full time under the new law.  If a business has 50 or more full-time employees, health care coverage has to be provided.  Except that a lot business owners say that the additional cost is going to be a financial killer, so instead, some of them have been cutting back hours to below that 30-hour threshold.  More than 200,000 Californians are at risk of losing hours from the health care law - that according to one study.

Julian: What kinds of businesses are doing this?

Lacter: Restaurant chains have received much of the attention, but the city of Long Beach, as an example, is going to reduce hours for a couple of hundred of its workers.  And, last week came word that the L.A.-based clothing chain Forever 21 will cut some of its full-time employees to a maximum 29-and-a-half hours a week, and classify them as part time.  That touched off an outcry on the Internet - people were saying that Forever 21 was being unfair and greedy - though the company says that only a small number of employees are affected, and that its decision has nothing to do with the Affordable Care Act.  There's really no way to know - Forever 21 is a private company, which means it's not obligated to disclose a whole lot.  What we do know is that those people will be losing their health care coverage.

Julian: And, the ultimate impact on businesses and workers?

Lacter: Steve, you're looking at several years before the picture becomes clear.  Here in California, workers not eligible for health care through their employer can get their own individual coverage, and if their income levels are not over a certain amount, they'd be eligible for Medicaid.  And, let's not forget many businesses already provide coverage for their employees.  So, lots of rhetoric - but, not many conclusions to draw from, which does make you wonder why so many business owners are unwilling to at least give this thing a chance.  Just doesn't seem to be much generosity of spirit for their workers, not to mention any recognition that if people can go to a doctor instead of an emergency room we'd probably all be better off.

Julian: Health care is far from the only controversy for Forever 21, true?

Lacter: In some ways, it's one of the biggest Southern California success stories.  Don Chang emigrated here in 1981 from Korea at the age of 18, opened his first store in Highland Park three years later (it was called Fashion 21), and he never looked back.  Today, revenues are approaching $4 billion.  But, the guy must have some pretty hefty legal bills because his company has been accused of all kinds of workplace violations.  The lawsuits alleged that workers preparing items for the Forever 21 stores didn't receive overtime, that they didn't get required work breaks, that they received substandard wages, and that they worked in dirty and unsafe conditions - sweatshop conditions, essentially.

Julian: Are most of their claims settled out of court?  You don't hear much about them.

Lacter: They are, which means there's usually a minimal amount of media coverage.  If a privately held company decides to keep quiet by not releasing financial results or other operational information, there's not likely to be much of a story - unlike what happens with a company like Apple, which is always under scrutiny.  Sometimes, plaintiffs will try to organize class-action suits, but that's extremely tough when you're dealing with low-wage workers who are often very reluctant to get involved because of their legal status.  And, let's not forget that Forever 21 - like any low-cost retailer - is simply catering to the demand for cheap, stylish clothes that are made as quickly as possible.

Julian: I guess you can't make that happen when wages and benefits are appreciably higher than your competition.

Lacter: The next time you walk into a Forever 21 store and wonder how prices can be so reasonable, that's how.

Mark Lacter writes for Los Angeles Magazine and pens the business blog at LA Observed.com.

This content is from Southern California Public Radio. View the original story at SCPR.org.




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