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Journal of Cell Science

Julie Milanini, Racha Fayad, Mariagrazia Partisani, Patrick Lecine, Jean-Paul Borg, Michel Franco, Frédéric Luton
A key step of epithelial morphogenesis is the creation of the lumen. Luminogenesis by hollowing proceeds through the fusion of apical vesicles at cell-cell contact. The small nascent lumens grow through extension, coalescence and enlargement coordinated with cell division to give rise to a single central lumen. Here, using MDCK cells grown in 3D-culture, we show that EFA6A participates in luminogenesis. EFA6A recruits α-actinin 1 (ACTN1) through direct binding. In polarized cells, ACTN1 was found to be enriched at the tight junction where it acts as a primary effector of EFA6A for normal luminogenesis...
December 15, 2017: Journal of Cell Science
Guillaume Hatte, Claude Prigent, Jean-Pierre Tassan
Epithelia are layers of polarised cells tightly bound to each other by adhesive contacts. Epithelia act as barriers between an organism and its external environment. Understanding how epithelia maintain their essential integrity while remaining sufficiently plastic to allow events such as cytokinesis to take place is a key biological problem. In vertebrates, the remodelling and reinforcement of adherens junctions maintains epithelial integrity during cytokinesis. The involvement of tight junctions in cell division, however, has remained unexplored...
December 15, 2017: Journal of Cell Science
Fidel Lozano-Elena, Ainoa Planas-Riverola, Josep Vilarrasa-Blasi, Rebecca Schwab, Ana I Caño-Delgado
Stem cell regeneration is crucial for both cell turnover and tissue healing in multicellular organisms. In Arabidopsis roots, a reduced group of cells known as the quiescent center (QC) acts as a cell reservoir for surrounding stem cells during both normal growth and in response to external damage. Although cells of the QC have a very low mitotic activity, plant hormones such as brassinosteroids (BR) can promote QC divisions. Here, we used a tissue-specific strategy to investigate the spatial signaling requirements of BR-mediated QC divisions...
December 14, 2017: Journal of Cell Science
Jefri Heyman, Balkan Canher, Anchal Bisht, Fien Christiaens, Lieven De Veylder
Plants react to wounding through the activation of both defense and repair pathways, but how these two responses are coordinated is unclear. Here, we put forward the hypothesis that diverse members of the subfamily X of the plant-specific ethylene response factor (ERF) transcription factors coordinate stress signaling with the activation of wound repair mechanisms. Moreover, we highlight the observation that tissue repair is strongly boosted through the formation of a heterodimeric protein complex that comprises ERF and transcription factors of the GRAS domain type...
December 14, 2017: Journal of Cell Science
Shunyao Xu, Zhen Zhou, Hao Li, Ziying Liu, Xiaojun Pan, Fen Wang, Yueyue Huang, Xiaogang Li, Yunbei Xiao, Jingye Pan, Cong Wang, Dequan Li
Sepsis is an aggressive and life-threatening systemic inflammatory response with a high mortality. Inflammation and coagulation play crucial roles in the pathogenesis of sepsis in a mutually promoting manner. Unlike other single-target molecular therapies that have no obvious effects on clinical sepsis, the bone marrow stromal cell (BMSC) therapy offers a broader spectrum of activities ranging from immune and inflammation suppression to tissue regeneration. In this report, we demonstrated that BMSC injection attenuated the septic coagulopathy...
December 14, 2017: Journal of Cell Science
Victoria G Shuttleworth, Luke Gaughan, Lotfia Nawafa, Caitlin A Mooney, Steven L Cobb, Neil S Sheerin, Ian R Logan
Chronic kidney disease (CKD) is a global socioeconomic problem. It is characterised by the presence of differentiated myofibroblasts that, in response to TGF B-1, produce tissue fibrosis, leading to renal failure. Here we define a novel interaction between the SET9 lysine methyltransferase and SMAD3, the principle mediator of TGF B-1 signalling in myofibroblasts. We show that SET9 deficient fibroblasts exhibit globally altered gene expression profiles in response to TGF B-1, whilst overexpression of SET9 enhances SMAD3 transcriptional activity...
December 8, 2017: Journal of Cell Science
M P Keasey, C Jia, L F Pimentel, R S Sante, C Lovins, T Hagg
We defined how blood-derived vitronectin (VTN) rapidly and potently activates leukemia inhibitory factor (LIF) and pro-inflammatory interleukin 6 (IL-6) in vitro and after vascular injury in the brain. VTN (not fibrinogen, fibronectin, laminin-111, collagen-I) robustly increased LIF and IL-6 within 4 hr in C6-astroglioma cells, while VTN-/- mouse plasma was less effective than wildtype. LIF and IL-6 were induced by intracerebral injection of rhVTN in mice, but less by intracerebral hemorrhage in VTN-/- than wildtype littermates...
December 8, 2017: Journal of Cell Science
Hilary Renshaw, José M Vargas-Muñiz, Praveen R Juvvadi, Amber D Richards, Greg Waitt, Erik J Soderblom, M Arthur Moseley, William J Steinbach
Myosins are critical motor proteins that contribute to the secretory pathway, polarized growth, and cytokinesis. The globular tail domains of class V myosins have been shown to be important for cargo binding and actin cable organization. Additionally, phosphorylation plays a role in class V myosin cargo choice. Our previous studies on the class V myosin, MyoE, in the fungal pathogen Aspergillus fumigatus confirmed its requirement for normal morphology and virulence. However, the domains and molecular mechanisms governing MyoE's function remain unknown...
December 8, 2017: Journal of Cell Science
Feike Hao, Kazuhiko Kondo, Takashi Itoh, Sumiko Ikari, Shigeyuki Nada, Masato Okada, Takeshi Noda
In response to amino acid supply, mTORC1, a master regulator of cell growth, is recruited to the lysosome and activated by a small GTPase, Rheb. However, the intracellular localization of Rheb is controversial. In this study, we showed that a significant portion of Rheb was localized on the Golgi but not on the lysosome. GFP-Rheb could activate mTORC1, even when forced to exclusively localize to the Golgi. Likewise, artificial recruitment of mTORC1 to the Golgi allowed its activation. Accordingly, the Golgi was in contact with the lysosome at the newly described Golgi lysosome contact site (GLCS)...
December 8, 2017: Journal of Cell Science
Sribalaji Lakshmikanthan, Magdalena Sobczak, Sergio Li Calzi, Lynn Shaw, Maria B Grant, Magdalena Chrzanowska-Wodnicka
Vascular Endothelial Growth Factor (VEGF), a key angiogenic and permeability factor, plays an important role in new blood vessel formation. However, abnormal VEGF-induced VEGFR2 signaling leads to hyper-permeability. We demonstrated that Rap1, best known for promoting cell adhesion and vessel stability, is a critical regulator of VEGFR2-mediated angiogenic and shear-stress EC responses. To determine Rap1's role in endothelial barrier dynamics, we examined vascular permeability in EC-specific Rap1A- and Rap1B- knockout mice, cell-cell junction remodeling and EC monolayer resistivity in Rap1-deficient ECs under basal, inflammatory or elevated VEGF conditions...
December 8, 2017: Journal of Cell Science
Christine Dolde, Joachim Bischof, Simon Grüter, Anna Montada, Jakob Halekotte, Christian Peifer, Hubert Kalbacher, Ulrich Baumann, Uwe Knippschild, Beat Suter
Casein kinase 1 (CK1) plays central roles in various signal transduction pathways and performs many cellular activities. For a long time CK1 was thought to act independent of modulatory subunits and in a constitutive manner. Recently, DEAD box RNA helicases, in particular DEAD box RNA helicase 3 X-linked (DDX3X), were found to stimulate CK1 activity in vitro In order to observe CK1 activity in living cells and to study its interaction with DDX3X, we developed a CK1-specific FRET biosensor. This tool revealed that DDX3X is indeed required for full CK1 activity in living cells...
December 8, 2017: Journal of Cell Science
Christina M Van Itallie, Amber Jean Tietgens, Angel Aponte, Marjan Gucek, Alexander X Cartagena-Rivera, Richard S Chadwick, James M Anderson
MARCKS-related protein (MRP) is tagged by biotin ligase fused to occludin and the level of tagging increases ∼20-fold following IFN/TNF treatment. GFP-MRP is focused at the lateral cell membrane and its overexpression potentiates the physiological response of the tight junction barrier to cytokines. However, deletion of MRP does not abrogate cytokine responses, suggesting that MRP is not required in the occludin-dependent IFN/TNF response. Instead, our results reveal a key role for MRP in epithelial cells in control of multiple actin-based structures, likely by regulation of integrin signaling...
December 8, 2017: Journal of Cell Science
Wataru Yamamoto, Suguru Wada, Makoto Nagano, Kaito Aoshima, Daria Elisabeth Siekhaus, Junko Y Toshima, Jiro Toshima
Clathrin-mediated endocytosis requires the coordinated assembly of various endocytic proteins and lipids at the plasma membrane. Accumulating evidence demonstrates a crucial role for phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) in endocytosis, but specific roles for PtdIns(4)P other than as the biosynthetic precursor of PtdIns(4,5)P2 have not been clarified. In this study we investigated the role of PtdIns(4)P or PtdIns(4,5)P2 in receptor-mediated endocytosis through the construction of temperature-sensitive (ts) mutants for the PI 4-kinases Stt4p and Pik1p and the PtdIns(4) 5-kinase Mss4p...
November 30, 2017: Journal of Cell Science
Patrícia A Simões, Ricardo Celestino, Ana X Carvalho, Reto Gassmann
In mitosis, the molecular motor dynein is recruited to kinetochores by the Rod-Zw10-Zwilch complex (RZZ) and Spindly to control spindle assembly checkpoint (SAC) signaling and microtubule attachment. How the ubiquitous dynein co-factors Lis1 and NudE/L contribute to these functions remains poorly understood. Here, we show that the C. elegans NudE/L homolog NUD-2 is dispensable for dynein- and LIS-1-dependent mitotic spindle assembly in the zygote. This facilitates functional characterization of kinetochore-localized NUD-2, which is recruited by the CENP-F-like proteins HCP-1/2 independently of RZZ-Spindly and dynein-LIS-1...
November 30, 2017: Journal of Cell Science
Faten Koraïchi, Rémi Gence, Catherine Bouchenot, Sarah Grosjean, Isabelle Lajoie-Mazenc, Gilles Favre, Stéphanie Cabantous
The human Ras superfamily of small GTPases controls essential cellular processes such as gene expression and cell proliferation. As their deregulation is widely associated with human cancer, small GTPases and their regulatory proteins have become increasingly attractive for the development of novel therapeutics. Classical methods to monitor GTPase activation include pulldown assays that limit the analysis of GTP-bound form of proteins from cell lysates. Alternatively, live-cell FRET biosensors may be used to study GTPase activation dynamics in response to stimuli, but these sensors often require further optimization for high-throughput applications...
November 30, 2017: Journal of Cell Science
Kosuke Shiraishi, Takahiro Hioki, Akari Habata, Hiroya Yurimoto, Yasuyoshi Sakai
The yeast high-osmolarity glycerol (HOG) pathway plays a central role in stress responses. It is activated by various stress stimuli including hyperosmotic stress, oxidative stress, high-temperature stress, and arsenite. Hog1, the critical MAP kinase of the pathway, localizes to the nucleus in response to high-osmolarity conditions, but otherwise little is known about its intracellular dynamics and regulation. Using the methylotrophic yeast Candida boidinii, we found that CbHog1-Venus formed intracellular dot structures after high-temperature stress, in a reversible manner...
November 28, 2017: Journal of Cell Science
Shweta Yadav, Rajan Thakur, Plamen Georgiev, Senthilkumar Deivasigamani, Harini K, Girish Ratnaparkhi, Padinjat Raghu
Phosphatidylinositol transfer proteins (PITPs) are essential regulators of PLC signalling. The PI transfer domain (PITPd) of multi-domain PITPs is reported to be sufficient for in vivo function questioning the relevance of other domains in the protein. In Drosophila photoreceptors, loss of RDGBα, a multi-domain PITP localized to membrane contact sites(MCS), results in multiple defects during PLC signalling. Here we report that the PITPd of RDGBα does not localize to MCS and fails to support function during strong PLC stimulation...
November 27, 2017: Journal of Cell Science
Jeroen Bakker, Menno Spits, Jacques Neefjes, Ilana Berlin
When cell surface receptors engage their cognate ligands in the extracellular space, they become competent to transmit potent signals to the inside of the cell, thereby instigating growth, differentiation, motility and many other processes. In order to control these signals, activated receptors are endocytosed and thoroughly curated by the endosomal network of intracellular vesicles and proteolytic organelles. In this Review, we follow the epidermal growth factor (EGF) receptor (EGFR) from ligand engagement, through its voyage on endosomes and, ultimately, to its destruction in the lysosome...
November 27, 2017: Journal of Cell Science
Alexander Buffone, Nicholas R Anderson, Daniel A Hammer
The recruitment of immune cells during inflammation is regulated by a multi-step cascade of cell rolling, activation, adhesion, and transmigration through the endothelial barrier. Similarly, Hematopoietic Stem and Progenitor Cells (HSPCs) use this pathway to migrate and home to the bone marrow. After selectin-mediated braking, HSPCs migrate along the vascular endothelium on adhesion ligands including ICAM-1, VCAM-1 or MAdCAM-1. Here we report that both the KG1a stem cell line and primary bone marrow CD34+ HSPCs can migrate against the direction of fluid flow on surfaces coated with cell adhesion molecules (CAMs), a behavior thus far only reported in T lymphocytes...
November 27, 2017: Journal of Cell Science
Fan Tu, Jakub Sedzinski, Yun Ma, Edward M Marcotte, John B Wallingford
Multiciliated cells (MCCs) drive fluid flow in diverse tubular organs and are essential for the development and homeostasis of the vertebrate central nervous system, airway, and reproductive tracts. These cells are characterized by dozens or hundreds of motile cilia that beat in a coordinated and polarized manner. In recent years, genomic studies have not only elucidated the transcriptional hierarchy for MCC specification, but also identified myriad new proteins that govern MCC ciliogenesis, cilia beating, or cilia polarization...
November 27, 2017: Journal of Cell Science
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