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Journal of Cell Science

Ali Vural, Ersin Fadillioglu, Fatih Kelesoglu, Dzwokai Ma, Stephen M Lanier
Activator of G-protein Signaling 3 (AGS3) exhibits broad functional diversity and oscillates among different subcellular compartments in a regulated manner. AGS3 consists of a tetratricopeptide repeat (TPR) domain and a G-protein regulatory (GPR) domain. We tested the hypothesis that phosphorylation of the AGS3 GPR domain regulated its subcellular distribution and functionality. In contrast to the cortical and/or diffuse non-homogeneous distribution of WT AGS3. AGS3 lacking 24 S/T phosphorylation sites in the GPR domain localized to cytosolic puncta and this was dependent upon a single amino acid (T602)...
November 9, 2018: Journal of Cell Science
Vasundhara Kandachar, Beatrice M Tam, Orson L Moritz, Dusanka Deretic
The rhodopsin-VxPx-Arf4 complex initiates expansion of vertebrate rod photoreceptor cilia-derived light-sensing organelles through stepwise assembly of the conserved trafficking network. Here, we examine its role in the sorting of VAMP7/TI-VAMP-an R-SNARE possessing a regulatory longin domain (LD)-into rhodopsin transport carriers (RTCs). During RTC formation and trafficking, VAMP7 co-localizes with the ciliary cargo rhodopsin and interacts with the Rab11-Rabin8-Rab8 trafficking module. Rab11 and Rab8 bind VAMP7 LD, whereas Rabin8 interacts with the SNARE domain...
November 7, 2018: Journal of Cell Science
Tomoaki Nagai, Sachiho Mukoyama, Harumi Kagiwada, Naoki Goshima, Kensaku Mizuno
Primary cilia are antenna-like sensory organelles that transmit various extracellular signals. Ciliogenesis requires the removal of CP110 and its interactor CEP97 from the mother centriole for initiating ciliary axoneme extension, but the underlying mechanism remains unknown. Here we show that CEP97 is partially degraded upon serum starvation by the ubiquitin-proteasome system. CEP97 was polyubiquitinated in serum-starved cells, and overexpression of a non-ubiquitinatable CEP97 mutant effectively blocked CP110 removal and ciliogenesis induced by serum-starvation...
November 7, 2018: Journal of Cell Science
Yuki Tanabe, Yoichiro Kamimura, Masahiro Ueda
In eukaryotic chemotaxis, parallel signaling pathways regulate the spatiotemporal pseudopod dynamics at the leading edge of a motile cell through characteristic dynamics of an excitable system; however, differences in the excitability and the physiological roles of individual pathways remain to be elucidated. Here we found that two different pathways, soluble guanylyl cyclase (sGC) and phosphatidylinositol 3-kinase (PI3K), exhibited similar all-or-none responses but different refractory periods by simultaneous observations of their excitable properties...
November 7, 2018: Journal of Cell Science
Yuqing Xia, Ning Huang, Zhiquan Chen, Fangyuan Li, Guiliang Fan, Dandan Ma, Jianguo Chen, Junlin Teng
The proteinaceous linker of centrosomes is an important structure that allows the centrosome to function as a single microtubule organizing center (MTOC) of cells in interphase. However, the assembly mechanism of the centrosome linker components remains largely unknown. In this study, we identify CCDC102B as a new centrosome linker protein that is required for maintaining centrosome cohesion. CCDC102B is recruited to the centrosome by C-Nap1 and interacts with the centrosome linker components Rootletin and LRRC45...
November 7, 2018: Journal of Cell Science
Manuela Lavorato, Emanuele Loro, Valentina Debattisti, Tejvir Khurana, Clara Franzini-Armstrong
Mitochondria respond to stress and undergo fusion and fission at variable rates, depending on the cell status. To understand mitochondria behavior during muscle fatigue, we investigated mitochondria ultrastructure and the level of a fission and stress-related protein in fast muscle fibers of mice subjected to fatigue. Mice were forced running at increasing speed till exhaustion at 45'-1 hr. In other mice high-intensity muscle stimulation through the sciatic nerve simulated the forced treadmill exercise. We detect the unusual presence of a rare phenotype characterized by Elongated Mitochondria Constrictions (EMCs) connecting two separate segments of the original organelles...
November 7, 2018: Journal of Cell Science
Xiangduo Kong, Gladys Mae Saquilabon Cruz, Sally Loyal Trinh, Xu-Dong Zhu, Michael W Berns, Kyoko Yokomori
TRF2 binds to telomeric repeats and is critical for telomere integrity. Evidence suggests that it also localizes to non-telomeric DNA damage sites. However, this recruitment appears to be precarious and functionally controversial. We find that TRF2 recruitment to damage sites occurs by a two-step mechanism: the initial rapid recruitment (phase I) and stable and prolonged association with damage sites (phase II). Phase I is poly(ADP-ribose) polymerase (PARP)-dependent and requires the N-terminal basic domain...
November 7, 2018: Journal of Cell Science
Zhe-Long Jin, Yu-Jin Jo, Suk Namgoong, Nam-Hyung Kim
Dynamic reorganization of the actin cytoskeleton is fundamental to a number of cellular events, and various actin-regulatory proteins modulate actin polymerization and depolymerization. Cyclase-associated proteins (CAPs), highly conserved actin monomer-binding proteins, have been known to promote actin disassembly by enhancing the actin-severing activity of ADF/cofilin. In this study, we found that CAP1 regulated actin remodeling during mouse oocyte maturation. Efficient actin disassembly during oocyte maturation is essential for asymmetric division and cytokinesis...
November 7, 2018: Journal of Cell Science
Claudio Bussi, Javier M Peralta Ramos, Daniela S Arroyo, Jose I Gallea, Paolo Ronchi, Androniki Kolovou, Ji M Wang, Oliver Florey, Maria S Celej, Yannick Schwab, Nicholas T Ktistakis, Pablo Iribarren
Autophagic dysfunction and protein aggregation have been linked to several neurodegenerative disorders, but the exact mechanisms and causal connections are not clear and most work was done in neurons and not in microglial cells. Here we report that exogenous fibrillar but not monomeric alpha-synuclein (AS) induces autophagy in microglial cells. We extensively studied the dynamics of this response by both live-cell imaging and correlative light-electron microscopy (CLEM) and found that it correlates with lysosomal damage and is characterised by the recruitment of the selective autophagy-associated proteins TANK-binding kinase 1 (TBK1) and Optineurin (OPTN) to ubiquitinated lysosomes...
November 7, 2018: Journal of Cell Science
Afsoon Saadin, Michelle Starz-Gaiano
How vesicle trafficking components actively contribute to regulation of paracrine signaling is unclear. We genetically uncovered a requirement for α-Soluble NSF Attachment Protein (α-Snap) in the activation of the Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) pathway during Drosophila egg development. α-Snap, a well-conserved vesicle trafficking regulator, mediates association of N-ethylmaleimide-Sensitive Factor (NSF) and SNAREs to promote vesicle fusion. Depletion of α-Snap or the SNARE family member Syntaxin1A in epithelia blocks polar cells maintenance and prevents specification of motile border cells...
November 7, 2018: Journal of Cell Science
James F E Grey, Alastair Campbell-Ritchie, Nicola M Everitt, Alexander J Fezovich, Sally P Wheatley
Since the establishment of cell culture, common practice has been to grow adherent cells in 2D monolayers. Although cells behave completely differently when grown in these artificial conditions, the ease of 2D culturing has meant that this practice still prevails today, and adopting conditions that more closely reflect the natural microenvironment has been met with substantial inertia. The alternative, animal models that mimic natural human physiology, are less accessible, strictly regulated and require licences and expensive facilities...
November 7, 2018: Journal of Cell Science
Chien-Chang Huang, Tai-Yu Chiu, Tzu-Ying Lee, Hsin-Jui Hsieh, Chung-Chih Lin, Lung-Sen Kao
α-Synuclein is associated with Parkinson's disease. α-Synuclein is mainly localized in presynaptic terminals and regulates exocytosis, but its physiological roles remain controversial. We studied effects of soluble and aggregated α-synuclein on exocytosis and explored the molecular mechanism by which α-synuclein interacts with regulatory proteins, including Rab3A, Munc13-1 and Munc18-1, in order to regulate exocytosis. Using fluorescence recovery after photobleaching, overexpressed α-synuclein in PC12 cells was found to be in a monomeric form, which promotes exocytosis...
November 7, 2018: Journal of Cell Science
Caroline Barisch, Vera Kalinina, Louise H Lefrançois, Joddy Appiah, Ana T López-Jiménez, Thierry Soldati
Professional phagocytes have developed an extensive repertoire of autonomous immunity strategies to ensure killing of bacteria. Besides phagosome acidification and the generation of reactive oxygen species, deprivation of nutrients and the lumenal accumulation of toxic metals are essential to kill ingested bacteria or inhibit growth of intracellular pathogens. We use the soil amoeba Dictyostelium discoideum , a professional phagocyte that digests bacteria for nutritional purposes, to decipher the role of zinc poisoning during phagocytosis of non-pathogenic bacteria and visualize the temporal and spatial dynamics of compartmentalized, free zinc using fluorescent probes...
November 7, 2018: Journal of Cell Science
Ahmed Elbediwy, Hannah Vanyai, Maria-Del-Carmen Diaz-de-la-Loza, David Frith, Ambrosius P Snijders, Barry J Thompson
Human cells can sense mechanical stress acting upon Integrin adhesions and respond by sending the YAP (YAP1) and TAZ (WWTR1) transcriptional co-activators to the nucleus to drive TEAD-dependent transcription of target genes. How Integrin signaling activates YAP remains unclear. Here we identify a key role for the Enigma (PDLIM7) and Enigma-like (PDLIM5) family of PDZ and LIM domain containing proteins in Integrin-mediated mechanotransduction. YAP binds to PDLIM5/7 via its C-terminal PDZ binding motif (PBM), which is essential for full nuclear localization and activity of YAP...
November 7, 2018: Journal of Cell Science
Judy K VanSlyke, Bruce A Boswell, Linda S Musil
Lens epithelial cells are bound to the lens extracellular matrix capsule, a major component of which is laminin. After cataract surgery, surviving lens epithelial cells are exposed to increased levels of fibronectin. We asked if fibronectin influences lens cell fate using a serum-free primary lens epithelial cell culture system (DCDMLs). We found that culturing DCDMLs with plasma-derived fibronectin upregulated canonical TGFβ signaling relative to cells plated on laminin. Fibronectin-exposed cultures also showed increased, TGFβ signaling-dependent differentiation into the two cell types responsible for posterior capsule opacification after cataract surgery, namely myofibroblasts and lens fiber cells...
November 7, 2018: Journal of Cell Science
Viktoria Wollrab, Julio M Belmonte, Lucia Baldauf, Maria Leptin, François Nédeléc, Gijsje H Koenderink
Cytoskeletal networks of actin filaments and myosin motors drive many dynamic cell processes. A key characteristic of these networks is their contractility. Despite intense experimental and theoretical efforts, it is not clear what mechanism favors network contraction over expansion. Recent work points to a dominant role for the nonlinear mechanical response of actin filaments, which can withstand stretching but buckle upon compression. Here we present an alternative mechanism. We study how interactions between actin and myosin-2 at the single filament level translate into contraction at the network scale by performing time-lapse imaging on reconstituted quasi-2D-networks mimicking the cell cortex...
November 7, 2018: Journal of Cell Science
Yihua Wang, Shan Zhong, Christopher J Schofield, Peter J Ratcliffe, Xin Lu
Hypoxia plays a critical role at cellular and physiological levels in all animals. The responses to chronic hypoxia are, at least substantially, orchestrated by activation of the hypoxia inducible transcription factors (HIFs), whose stability and subsequent transcriptional activation are regulated the by HIF hydroxylases. Factor inhibiting HIF (FIH), initially isolated as a HIFα interacting protein following a yeast two-hybrid screen, is an asparaginyl hydroxylase that negatively regulates transcriptional activation by HIF...
October 17, 2018: Journal of Cell Science
Christoph Niemietz, Lutz Fleischhauer, Vanessa Sandfort, Sarah Guttmann, Andree Zibert, Hartmut H-J Schmidt
Transthyretin (TTR)-related familial amyloid polyneuropathy (ATTR) results from aggregation and extracellular disposition of misfolded TTR variants. Growing evidence suggests the importance of hepatic chaperones for modulation of pathogenesis. We took advantage of iPSC-derived hepatocyte-like cells (HLCs) derived from ATTR patients (ATTR-HLCs) to compare chaperone gene expression to healthy individuals (H-HLCs). From the set of genes analyzed, chaperones that are predominantly located extracellularly were differently expressed...
October 17, 2018: Journal of Cell Science
Catherine Cheng, Roberta B Nowak, Michael B Amadeo, Sondip K Biswas, Woo-Kuen Lo, Velia M Fowler
Tropomyosins (Tpms) stabilize F-actin and regulate interactions with other actin-binding proteins. The eye lens changes shape in order to fine focus light to transmit a clear image, and thus lens organ function is tied to its biomechanical properties, presenting an opportunity to study Tpm functions in tissue mechanics. Mouse lenses contain Tpm3.5 (TM5NM5), a previously unstudied isoform associated with F-actin on lens fiber cell membranes. Decreased levels of Tpm3.5 lead to softer and less mechanically resilient lenses that are unable to resume their original shape after compression...
October 17, 2018: Journal of Cell Science
Nicholas Mikolajewicz, Ali Mohammed, Martin Morris, Svetlana V Komarova
Body tissues are exposed to a complex mechanical environment which is perceived by cells and converted to biochemical signals, such as ATP release. We performed a meta-analysis of 278 systematically identified studies that investigated mechanically-stimulated ATP release (MSAR) to quantify the amounts, kinetics and mechanisms of ATP release under normal and pathological conditions. Mechanically-stimulated mammalian cells were shown to release 38.6 (95% CI: 18.2 to 81.8) amoles ATP/cell on average with a characteristic time constant of 32 s (95% CI: 16 to 66)...
October 17, 2018: Journal of Cell Science
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