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Journal of Cell Science

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https://www.readbyqxmd.com/read/29150488/membrane-binding-of-and-a-self-inhibitory-interaction-within-nup155-are-required-for-nuclear-pore-complex-formation
#1
Paola De Magistris, Marianna Tatarek-Nossol, Manfred Dewor, Wolfram Antonin
Nuclear pore complexes (NPCs) are the gateways through the nuclear envelope. How they form into a structure containing three rings and integrate into the nuclear envelope remains a challenging paradigm for coordinated assembly of macro-complexes. In vertebrates, the cytoplasmic and nucleoplasmic rings of NPCs are mostly formed by multiple copies of the Nup107-Nup160 complex, whereas the central, or inner ring is composed of Nup53, Nup93, Nup155 and the two paralogues Nup188 and Nup205. Inner ring assembly is only partially understood...
November 17, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29150487/dynamin-inhibitors-block-mtorc1-activation-by-amino-acids-independently-of-dynamin
#2
Persaud Avinash, Cormerais Yann, Pouyssegur Jacques, Rotin Daniela
mTORC1 plays a critical role in protein synthesis and cell proliferation and growth. It is activated by growth factors and amino acids, including essential amino acid (EAA), such as Leu; Leu enters cells via the Leu transporter LAT1-4F2hc and potentially via endocytosis. Here we investigated the contribution of the different routes of Leu entry into cells to mTORC1 activation using pharmacological inhibitors and cells that lack LAT1 or dynamin1/2/3. Our results show that LAT1 is the major route of Leu entry into cells and mTORC1 activation (∼70%), while dynamin-dependent endocytosis and macropinocytosis contribute minimally to both (5-15%)...
November 17, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29142103/an-intracellular-activation-of-smoothened-independent-of-hedgehog-stimulation-in-drosophila
#3
Kai Jiang, Yajuan Liu, Jie Zhang, Jianhang Jia
Smoothened (Smo), a GPCR family protein, plays a critical role in the reception and transduction of Hedgehog (Hh) signal. Smo is phosphorylated and activated on the cell surface, however, it is unknown whether Smo can be intracellularly activated. Here, we demonstrate that inactivation of the ESCRT-III causes dramatic accumulation of Smo, and subsequent activation of Hh signaling. In contrast, inactivation of ESCRTs 0-II induces mild Smo accumulation. We provide evidence that Kurtz (Krz), the Drosophila β-arrestin2, acts in parallel with the ESCRTs 0-II pathway to sort Smo to the multivesicular bodies and lysosome-mediated degradation...
November 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29142102/prolines-in-the-%C3%AE-helix-confer-the-structural-flexibility-and-functional-integrity-of-importin-%C3%AE
#4
Masahiro Kumeta, Hide A Konishi, Wanzhen Zhang, Sayuri Sakagami, Shige H Yoshimura
The karyopherin family of nuclear transport receptors is composed of a long array of amphiphilic α-helices and undergoes flexible conformational changes to pass through the hydrophobic crowding barrier of the nuclear pore. Here, we focused on the characteristic enrichment of prolines in the middle of the outer α-helices of importin β. When these prolines were substituted with alanine, nuclear transport activity was reduced drastically in vivo and in vitro, and caused a severe defect in mitotic progression...
November 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29142101/loss-of-adam9-expression-impairs-%C3%AE-1-integrin-endocytosis-focal-adhesion-formation-and-cancer-cell-migration
#5
Kasper J Mygind, Jeanette Schwarz, Pranshu Sahgal, Johanna Ivaska, Marie Kveiborg
The transmembrane protease ADAM9 is frequently upregulated in human cancers and it promotes tumour progression in mice. In vitro, ADAM9 regulates cancer cell adhesion and migration by interacting with integrins. Yet, how ADAM9 modulates integrin functions is not known. We here show that ADAM9 knockdown increases β1 integrin levels through mechanisms independent of its protease activity. In ADAM9-silenced cells, adhesion to collagen and fibronectin is reduced, suggesting an altered function of the accumulated integrins...
November 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29133588/quantitative-analysis-of-multilayer-organization-of-proteins-and-rna-in-nuclear-speckles-at-super-resolution
#6
Jingyi Fei, Mahdieh Jadaliha, Tyler S Harmon, Isaac T S Li, Boyang Hua, Qinyu Hao, Alex S Holehouse, Matthew Reyer, Qinyu Sun, Susan M Freier, Rohit V Pappu, Kannanganattu V Prasanth, Taekjip Ha
Nuclear speckles are self-assembled organelles composed of RNAs and proteins. They are proposed to act as structural domains that control distinct steps in gene expression, including transcription, splicing, and mRNA export. Earlier studies identified differential localization of a few components within the speckles. It was speculated that the spatial organization of speckle components might contribute directly to the order of operations that coordinate distinct processes. By performing multi-color structured illumination microscopy, we characterized the multilayer organization of speckles at a higher resolution...
November 13, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29122984/effects-of-mutating-%C3%AE-tubulin-lysine-40-on-sensory-dendrite-development
#7
Brian V Jenkins, Harriet A J Saunders, Helena L Record, Dena M Johnson-Schlitz, Jill Wildonger
Microtubules are essential to neuronal structure and function. Axonal and dendritic microtubules are enriched in post-translational modifications that impact microtubule dynamics, transport, and microtubule-associated proteins. Acetylation of α-tubulin lysine 40 (K40) is a prominent, conserved modification of neuronal microtubules. However, the cellular role of microtubule acetylation remains controversial. To resolve how microtubule acetylation might affect neuronal morphogenesis we mutated endogenous α-tubulin in vivo using a new fly strain that facilitates the rapid knock-in of designer α-tubulin alleles...
November 9, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29122983/restricted-cell-functions-on-micropillars-are-alleviated-by-surface-nanocoating-with-amino-groups
#8
Caroline Moerke, Susanne Staehlke, Henrike Rebl, Birgit Finke, J Barbara Nebe
The topographical and chemical surface features of biomaterials are sensed by the cells, affecting their physiology at the interface. On titanium we recently discovered an attempted caveolae-mediated phagocytosis of the sharp-edged microstructures by osteoblasts. This active, energy-consuming process resulted in decreased osteoblastic cell functions, e.g. extracellular matrix proteins. However, chemical modification with plasma polymerized allylamine (PPAAm) was able to amplify osteoblast adhesion and spreading, resulting in better implant osseointegration in vivo In the present in vitro study we analyzed whether this plasma polymer nanocoating is able to attenuate the microtopography-induced changes of osteoblast physiology...
November 9, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29122982/real-time-observation-of-light-controlled-transcription-in-living-cells
#9
Anne Rademacher, Fabian Erdel, Jorge Trojanowski, Sabrina Schumacher, Karsten Rippe
Gene expression is tightly regulated in space and time. To dissect this process with high temporal resolution, we introduce an optogenetic tool termed BLInCR (Blue Light-Induced Chromatin Recruitment) that combines rapid and reversible light-dependent recruitment of effector proteins with a real-time readout for transcription. We used BLInCR to control the activity of a reporter gene cluster in the human osteosarcoma cell line U2OS by reversibly recruiting the viral transactivator VP16. RNA production was detectable ∼2 minutes after VP16 recruitment and readily decreased when VP16 dissociated from the cluster in the absence of light...
November 9, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29122981/in-situ-imaging-of-mitochondrial-translation-shows-weak-correlation-with-nucleoid-dna-intensity-and-no-suppression-during-mitosis
#10
Christopher Estell, Emmanouela Stamatidou, Sarah El-Messeiry, Andrew Hamilton
Although mitochondrial translation produces only 13 proteins, we show here how this can be labelled and detected in situ by fluorescence microscopy with a simple, rapid and inexpensive procedure using non-canonical amino acid labelling and click chemistry. This allows visualisation of the translational output in different mitochondria within a cell, their position within that cell and a comparison of mitochondrial translation between cells. Most highly translationally active mitochondria were closest to the nucleus but were also found at the distal end of long cellular projections...
November 9, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29113998/rhoc-regulates-actin-remodeling-to-form-phagosomes-during-fc%C3%AE-r-mediated-phagocytosis
#11
Youhei Egami, Katsuhisa Kawai, Nobukazu Araki
Phagosome formation is a complicated process that requires spatiotemporally regulated actin reorganization. We found that RhoC GTPase is a critical regulator of FcγR-mediated phagocytosis in macrophages. Our live-cell imaging revealed that RhoC, but not RhoA, is recruited to phagocytic cups engulfing IgG-opsonized erythrocytes (IgG-Es). RhoC silencing by RNAi, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-mediated RhoC knockout and the expression of dominant-negative or dominant-active RhoC mutants suppressed the phagocytosis of IgG-Es...
November 7, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29113997/imaedge-a-platform-for-the-quantitative-analysis-of-cortical-proteins-spatiotemporal-dynamics-during-cell-polarization
#12
Zhen Zhang, Yen Wei Lim, Peng Zhao, Pakorn Kanchanawong, Fumio Motegi
Cell polarity involves the compartmentalization of the cell cortex. The establishment of cortical compartments arises from the spatial bias in the activity and concentration of cortical proteins. The mechanistic dissection of cell polarity requires the accurate detection of dynamic changes in cortical proteins, but the fluctuations of cell shape and the inhomogeneous distributions of cortical proteins greatly complicate the quantitative extraction of their global and local changes during cell polarization. To address these problems, we introduce an open-source software package, ImaEdge, which automates the segmentation of the cortex from time-lapse movies, and enables quantitative extraction of cortical protein intensities...
November 7, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29097383/mind-the-sr-gap-roles-of-slit-robo-gaps-in-neurons-brains-and-beyond
#13
REVIEW
Bethany Lucas, Jeff Hardin
The Slit-Robo GTPase-activating proteins (srGAPs) were first identified as potential Slit-Robo effectors that influence growth cone guidance. Given their N-terminal F-BAR, central GAP and C-terminal SH3 domains, srGAPs have the potential to affect membrane dynamics, Rho family GTPase activity and other binding partners. Recent research has clarified how srGAP family members act in distinct ways at the cell membrane, and has expanded our understanding of the roles of srGAPs in neuronal and non-neuronal cells...
November 2, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29093023/creb3l1-mediated-functional-and-structural-adaptation-of-the-secretory-pathway-in-hormone-stimulated-thyroid-cells
#14
Iris A García, Vanina Torres Demichelis, Diego L Viale, Pablo Di Giusto, Yulia Ezhova, Roman S Polishchuk, Luciana Sampieri, Hernán Martinez, Elizabeth Sztul, Cecilia Alvarez
Many secretory cells increase the synthesis and secretion of cargo proteins in response to specific stimuli. How cells couple increased cargo load with a coordinate rise in secretory capacity to ensure efficient transport is not well understood. We used thyroid cells stimulated with thyrotropin (TSH) to demonstrate a coordinate increase in the production of thyroid-specific cargo proteins and ER-Golgi transport factors, and a parallel expansion of the Golgi complex. TSH also increased expression of the CREB3L1 transcription factor, which alone caused amplified transport factor levels and Golgi enlargement...
November 1, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29093022/giantin-knockout-models-reveal-a-feedback-loop-between-golgi-function-and-glycosyltransferase-expression
#15
Nicola L Stevenson, Dylan J M Bergen, Roderick E H Skinner, Erika Kague, Elizabeth Martin-Silverstone, Kate A Robson Brown, Chrissy L Hammond, David J Stephens
The Golgi is the cellular hub for complex glycosylation, controlling accurate processing of complex proteoglycans, receptors, ligands, and glycolipids. Its structure and organisation is dependent on golgins, which tether cisternal membranes and incoming transport vesicles. Here we show that knockout of the largest golgin, giantin, leads to substantial changes in gene expression despite only limited effects on Golgi structure. Notably, 22 Golgi-resident glycosyltransferases, but not glycan processing enzymes or the ER glycosylation machinery, are differentially expressed following giantin ablation...
November 1, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29084823/phosphorylation-of-the-rna-binding-protein-zfs1-modulates-sexual-differentiation-in-fission-yeast
#16
Francisco J Navarro, Probir Chakravarty, Paul Nurse
Sexual differentiation in the fission yeast Schizosaccharomyces pombe promotes cell cycle arrest and extensive changes in gene expression, resulting in cell-to-cell fusion, the exchange of hereditary material, and specialized cell division. These events are detrimental to the cell if they are triggered in inappropriate conditions, and therefore the decision to differentiate must be precisely controlled. Here, we investigated the role of the RNA-binding protein Zfs1 in this process by identifying its targets and characterizing novel posttranslational regulation...
October 30, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29084822/dynamic-changes-in-the-interchromosomal-interaction-of-early-histone-gene-loci-during-early-development-of-sea-urchin
#17
Masaya Matsushita, Hiroshi Ochiai, Ken-Ichi T Suzuki, Sayaka Hayashi, Takashi Yamamoto, Akinori Awazu, Naoaki Sakamoto
The nuclear positioning and chromatin dynamics of eukaryotic genes are closely related to the regulation of gene expression, but they have not been well examined during early development, which is accompanied by rapid cell cycle progression and dynamic changes in nuclear organization, such as nuclear size and chromatin constitution. In this study, we focused on the early development of the sea urchin Hemicentrotus pulcherrimus and performed three-dimensional fluorescence in situ hybridization of early histone gene loci...
October 30, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29074579/phragmoplast-microtubule-dynamics-a-game-of-zones
#18
REVIEW
Andrei Smertenko, Seanna L Hewitt, Caitlin N Jacques, Rafal Kacprzyk, Yan Liu, Matthew J Marcec, Lindani Moyo, Aaron Ogden, Hui Min Oung, Sharol Schmidt, Erika A Serrano-Romero
Plant morphogenesis relies on the accurate positioning of the partition (cell plate) between dividing cells during cytokinesis. The cell plate is synthetized by a specialized structure called the phragmoplast, which consists of microtubules, actin filaments, membrane compartments and associated proteins. The phragmoplast forms between daughter nuclei during the transition from anaphase to telophase. As cells are commonly larger than the originally formed phragmoplast, the construction of the cell plate requires phragmoplast expansion...
October 26, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29074578/dynein-and-eff-1-control-dendrite-morphology-through-regulating-the-localization-pattern-of-sax-7-in-epidermal-cells
#19
Ting Zhu, Xing Liang, Xiangming Wang, Kang Shen
Our previous work showed the cell adhesion molecule SAX-7 formed an elaborate pattern in epidermal cell, which instructs PVD dendrite branching. However, the molecular mechanism forming the SAX-7 pattern in the epidermis is not fully understood. Here, we report that the dynein light intermediate chain DLI-1, and the fusogen EFF-1 are required in epidermal cells to pattern SAX-7. While previous reports suggest these two molecules act cell-autonomously in PVD, our results showed the disorganized PVD dendritic arbors in these mutants are due to the abnormal SAX-7 localization patterns in epidermal cells...
October 26, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29074577/functional-analyses-of-the-cif1-cif2-complex-in-trypanosome-identify-the-structural-motifs-required-for-cytokinesis
#20
Huiqing Hu, Paul Majneri, Dielan Li, Yasuhiro Kurasawa, Tai An, Gang Dong, Ziyin Li
Cytokinesis in trypanosome occurs uni-directionally along the longitudinal axis from the cell anterior towards the cell posterior and requires a trypanosome-specific CIF1-CIF2 protein complex. However, little is known about the contribution of the structural motifs in CIF1 and CIF2 to complex assembly and cytokinesis. Here, we demonstrated that the two zinc-finger motifs but not the coiled-coil motif in CIF1 are required for interaction with the EF-hand motifs in CIF2. We further showed that localization of CIF1 depends on the coiled-coil motif and the first zinc-finger motif and that localization of CIF2 depends on the EF-hand motifs...
October 26, 2017: Journal of Cell Science
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