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Journal of Cell Science

Dan Zhao, Xiao-Man Liu, Zhong-Qiu Yu, Ling-Ling Sun, Xingchuang Xiong, Meng-Qiu Dong, Li-Lin Du
Autophagy cargos include not only soluble cytosolic materials but also bulky organelles such as ER and mitochondria. In budding yeast, two proteins that contain the PX domain and the BAR domain, Atg20/Snx42 and Atg24/Snx4, are required for organelle autophagy and contribute to general autophagy in a way that can be compensated. It remains unclear why these proteins are important for organelle autophagy. Here, we show that in a distantly related fungal organism the fission yeast Schizosaccharomyces pombe, autophagy of ER and mitochondria is induced by nitrogen starvation and is promoted by three Atg20 and Atg24 family proteins, Atg20, Atg24, and Atg24b/SPBC1711...
October 13, 2016: Journal of Cell Science
Aidan P Maartens, Jutta Wellmann, Emma Wictome, Benjamin Klapholz, Hannah Green, Nicholas H Brown
Vinculin is a highly conserved protein involved in cell adhesion and mechanotransduction, and both gain and loss of its activity causes defective cell behaviour. Here, we examine how altering vinculin activity perturbs integrin function within the context of Drosophila development. Whereas loss of vinculin produced relatively minor phenotypes, gain of vinculin activity, via a loss of head-tail autoinhibition, caused lethality. The minimal domain capable of inducing lethality is the talin-binding D1 domain, and this appears to require talin-binding activity, as lethality was suppressed by competition with single vinculin binding sites from talin...
October 13, 2016: Journal of Cell Science
John Maringa Githaka, Anthony R Vega, Michelle A Baird, Michael W Davidson, Khuloud Jaqaman, Nicolas Touret
Nanoclustering is an emerging organizational principle of membrane-associated proteins. The functional consequences of nanoclustering for receptor signaling remain largely unknown. Here we applied quantitative, multi-channel high- and super-resolution imaging to the endothelial cell surface receptor CD36, the clustering of which upon binding to multivalent ligands such as the anti-angiogenic factor thrombospondin-1 (TSP-1) is thought to be critical for signaling. We found that a substantial fraction of unligated CD36 exists in nanoclusters which not only promote TSP-1 binding but are also enriched with the downstream effector Fyn...
September 30, 2016: Journal of Cell Science
Takashi Okabe, Rohit Chavan, Sara S Fonseca Costa, Andrea Brenna, Jürgen A Ripperger, Urs Albrecht
REV-ERBα (Nr1d1) is a nuclear receptor that is part of the circadian clock mechanism and regulates metabolism and inflammatory processes. The glucocorticoid receptor (GR, Nr3c1) influences similar processes, but is not part of the circadian clock although glucocorticoid signaling affects resetting of the circadian clock in peripheral tissues. Because of their similar impact on physiological processes we studied the interplay between these two nuclear receptors. We found that REV-ERBα binds to the C-terminal part and GR to the N-terminal part of HSP90, a chaperone responsible for the activation of proteins to ensure survival of a cell...
September 29, 2016: Journal of Cell Science
Evanthia Pangou, Christina Befani, Ilias Mylonis, Martina Samiotaki, George Panayotou, George Simos, Panagiotis Liakos
Hypoxia Inducible Factor 2 (HIF-2) is a transcriptional activator implicated in the cellular response to hypoxia. Regulation of its inducible subunit, HIF-2α, involves post-translational modifications. Here, we demonstrate that casein kinase 1δ (CK1δ) phosphorylates HIF-2α at Ser383 and Thr528 in vitro Disruption of these phosphorylation sites and silencing or chemical inhibition of CK1δ reduced the expression of HIF-2 target genes and the secretion of erythropoietin (EPO) in two hepatic cancer cell lines, Huh7 and HepG2, without affecting levels of HIF-2α protein expression...
September 29, 2016: Journal of Cell Science
Daniel Effelsberg, Luis Daniel Cruz-Zaragoza, Wolfgang Schliebs, Ralf Erdmann
Peroxisomal proteins carrying a type 1 peroxisomal targeting signal (PTS1) are recognized by the well-conserved cycling import receptor Pex5p. Yeast YMR018w gene codes for a Pex5p paralog and novel peroxin that is involved in peroxisomal import of a subset of matrix proteins. The novel peroxin was designated Pex9p and it interacts with the docking protein Pex14p and a subclass of PTS1-containing peroxisomal matrix enzymes. Unlike Pex5p, Pex9p is not expressed in glucose- or ethanol-grown cells, but it is strongly induced by oleate...
September 27, 2016: Journal of Cell Science
Xingyu Zhou, Mingsen Li, Huaxing Huang, Keren Chen, Zhuning Yuan, Ying Zhang, Yaping Nie, Hu Chen, Xumeng Zhang, Luxi Chen, Yaosheng Chen, Delin Mo
Although the mechanism underlying modulation of transcription factors in myogenesis has been well elucidated, the function of the transcription cofactors involved in this process remains poorly understood. Here, we identified HMGB2 as an essential nuclear transcriptional co-regulator in myogenesis. HMGB2 was highly expressed in undifferentiated myoblasts and regenerating muscle. Knockdown of HMGB2 inhibited myoblast proliferation and stimulated its differentiation. HMGB2 depletion down-regulated Myf5 and Cyclin A2 on the protein but not mRNA level...
September 26, 2016: Journal of Cell Science
Angelika Riedl, Michaela Schlederer, Karoline Pudelko, Mira Stadler, Stefanie Walter, Daniela Unterleuthner, Christine Unger, Nina Kramer, Markus Hengstschläger, Lukas Kenner, Dagmar Pfeiffer, Georg Krupitza, Helmut Dolznig
3D cancer models are used as preclinical systems to mimic physiologic drug response. We provide evidence for robust changes of proliferation and metabolic capacity in 3D by systematically analyzing spheroids of colon cancer cell lines. Spheroids showed relative lower AKT/mTOR/S6K activities compared to cells cultured in 2D. We identified spatial alterations in signaling, as the level of phospho-rpS6 decreased from the spheroid surface to the center, closely recapitulating the tumor areas around vessels in vivo These 3D-models displayed augmented anti-tumor response to AKT/mTOR/S6K- or MAPK-pathway inhibition compared to 2D...
September 23, 2016: Journal of Cell Science
Eden Yifrach, Silvia G Chuartzman, Noa Dahan, Shiran Maskit, Lior Zada, Uri Weill, Ido Yofe, Tsviya Olender, Maya Schuldiner, Einat Zalckvar
To optimally perform the diversity of metabolic functions that occur within peroxisomes, cells must dynamically regulate peroxisome size, number and content in response to cell state and the environment. Except for transcriptional regulation little is known about the mechanisms used to perform this complicated fete. Here we used complementary high content screens to follow changes in most of the yeast, Saccharomyces cerevisiae, proteins during growth in oleate. We found extensive changes in cellular architecture and identified several proteins co-localizing with peroxisomes that have not previously been considered peroxisomal proteins...
September 23, 2016: Journal of Cell Science
Linzhang Li, Jie Chen, Gaofeng Xiong, Daret K St Clair, Wei Xu, Ren Xu
Loss of epithelial cell polarity promotes cell invasion and cancer dissemination. Therefore, identification of factors that disrupt polarized acinar formation is critical. Reactive oxygen species (ROS) drive cancer progression and promote inflammation. Here, we show that the non-polarized breast cancer cell line, T4-2, generates significantly higher ROS levels than polarized S1 and T4R cells in 3D (three dimensional) culture, accompanied by induction of the nuclear factor κB (NF-κB) pathway and cytokine expression...
September 21, 2016: Journal of Cell Science
Emma L Bastow, Amber R Peswani, Daniel S J Tarrant, Daniel R Pentland, Xi Chen, Alan Morgan, Gemma L Staniforth, Jennifer M Tullet, Michelle L Rowe, Mark J Howard, Mick F Tuite, Campbell W Gourlay
A number of genes have been linked to familial forms of the fatal motor neuron disease Amyotrophic Lateral Sclerosis (ALS). Over 150 mutations within the SOD1 gene have been implicated in ALS, but why such mutations lead to ALS-associated cellular dysfunction is unclear. In this study, we identify how ALS-linked SOD1 mutations lead to changes in the cellular health of the yeast Saccharomyces cerevisiae We find that it is not the accumulation of aggregates, but instead the loss of Sod1 stability that drives cellular dysfunction...
September 21, 2016: Journal of Cell Science
Brad McColl, Ritu Garg, Philippe Riou, Kirsi Riento, Anne J Ridley
Rnd proteins are atypical members of the Rho GTPase family that induce actin cytoskeletal reorganization and cell rounding. Rnd proteins have been reported to bind to the intracellular domain of several Plexin receptors, but whether Plexins contribute to the Rnd-induced rounding response is not known. Here we show that Rnd3 interacts preferentially with Plexin-B2 of the three Plexin-B proteins, whereas Rnd2 interacts with all three B-type Plexins, and Rnd1 shows only very weak interaction with Plexin-B proteins in immunoprecipitations...
September 21, 2016: Journal of Cell Science
Yan Huang, Sheng Huang, Sin Man Lam, Zhihua Liu, Guanghou Shui, Yong Q Zhang
Nervous system development and function are tightly regulated by metabolic processes, including the metabolism of lipids such as fatty acids (FAs). Mutations in long-chain acyl-CoA synthetase 4 (ACSL4) are associated with non-syndromic intellectual disabilities. We previously reported that Acsl, the Drosophila ortholog of mammalian ACSL3 and ACSL4, inhibits neuromuscular synapse growth by suppressing transforming growth factor-β/bone morphogenetic protein (BMP) signaling. Here, we report that Acsl regulates the composition of FAs and membrane lipid, which in turn affect neuromuscular junction (NMJ) synapse development...
September 21, 2016: Journal of Cell Science
Dimitris Basagiannis, Sofia Zografou, Carol Murphy, Theodore Fotsis, Lucia Morbidelli, Marina Ziche, Christopher Bleck, Jason Mercer, Savvas Christoforidis
Endocytosis plays critical role in receptor signalling. VEGFR2 and its ligand VEGFA are fundamental in neovascularization. Yet, our understanding of the role of endocytosis in VEGFR2 signalling remains limited. Despite the existence of diverse internalisation routes, the only known endocytic pathway of VEGFR2 is the clathrin-mediated. Here, we show that this pathway is the predominant internalisation route of VEGFR2 only in the absence of ligand. Intriguingly, VEGF introduces a novel internalisation itinerary for VEGFR2, the pathway of macropinocytosis, which becomes the prevalent endocytic route of the receptor in the presence of ligand, while the route of clathrin becomes minor...
September 21, 2016: Journal of Cell Science
Pablo Luján, Giulia Varsano, Teresa Rubio, Marco L Hennrich, Timo Sachsenheimer, Manuel Gálvez-Santisteban, Fernando Martín-Belmonte, Anne-Claude Gavin, Britta Brügger, Maja Köhn
Disruption of epithelial architecture is a fundamental event during epithelial tumorigenesis. We show that the expression of the cancer-promoting phosphatase PRL-3 (PTP4A3), which is overexpressed in several epithelial cancers, in polarized epithelial MDCK and Caco2 cells leads to invasive and also multiple ectopic fully polarized lumen-containing cysts. Both processes disrupt epithelial architecture and are hallmarks of cancer. The pathological relevance of these findings is supported by the knock-down of endogenous PRL-3 in MCF-7 breast cancer cells grown in 3-dimensional branched structures, showing the rescue from multiple-lumen to single lumen containing branch ends...
September 21, 2016: Journal of Cell Science
Jonathan Bergeman, Alexia Caillier, François Houle, Laurence M Gagné, Marc-Étienne Huot
By progressing through the epithelial to mesenchymal transition (EMT), cancer cells gain the ability to leave the primary tumor site and invade surrounding tissues. These metastatic cancers cells can further increase their plasticity by adopting an amoeboid-like morphology, by undergoing mesenchymal to amoeboid transition (MAT). We found that adhering cells producing spreading initiation centers (SIC), a transient structure localized above nascent adhesion complexes, share common biological and morphological characteristics associated with amoeboid cells...
September 16, 2016: Journal of Cell Science
Tobin Stephanie Wales, Dabo Yang, Girgis John, Farahzad Ali, Alexandre Blais, John Charles McDermott
MEF2 and AP-1 transcription complexes have been individually implicated in myogenesis but their genetic interaction has not previously been addressed. Using MEF2A, c-Jun and Fra-1 ChIP-seq data and predicted AP-1 consensus motifs, we identified putative common MEF2 and AP-1 target genes, several of which are implicated in regulating the actin cytoskeleton. Since muscle atrophy results in remodelling or degradation of the actin cytoskeleton, we characterized the expression of putative MEF2/AP-1 target genes (Dstn, Flnc, Hspb7, Lmod3 and Plekhh2) under atrophic conditions using Dexamethasone (Dex) treatment in skeletal myoblasts...
September 15, 2016: Journal of Cell Science
Santiago Balseiro-Gomez, Juan A Flores, Jorge Acosta, M Pilar Ramirez-Ponce, Eva Ales
To ensure normal immune function, mast cells (MC) employ different pathways to release mediators. Here, we report a thus far unknown capacity of MC to recycle and reuse secretory granules (SG) after an antigen-evoked degranulation process under physiological conditions; this phenomenon involves the existence of a recycling SG pool available for release in a short time-scale. Approximately 60% of the total recycled SG was promoted by rapid endocytic modes, which involved kiss-and-run and cavicapture mechanisms, causing retention of the intragranular matrix...
September 13, 2016: Journal of Cell Science
Claire Heride, Daniel J Rigden, Erithelgi Bertsoulaki, Danilo Cucchi, Enrico De Smaele, Michael J Clague, Sylvie Urbé
USP21 is a centrosome-associated deubiquitylase (DUB) that has been implicated in the formation of primary cilia, critical organelles for the regulation of the Hedgehog (Hh) signaling pathway in vertebrates. Here we identify KCTD6, a Cullin3 E3-ligase substrate adapter previously linked to Hh-signaling, as well as Gli1, the key transcription factor responsible for Hh signal amplification, as novel interacting partners of USP21. We identify a cryptic structured protein interaction domain in KCTD6, which is predicted to bear fold similarity to Smr domains...
September 12, 2016: Journal of Cell Science
Wei Zhang, Amanda C Vreeland, Noa Noy
The RNA-binding protein HuR binds to AU-rich elements in target mRNAs and stabilizes them against degradation. The complete spectrum of genes whose expression is regulated by HuR and thus the basis for the broad range of cellular functions of the protein are incompletely understood. We show that HuR controls the expression of multiple components of the nuclear import machinery. Consequently, HuR is critical for the nuclear import of cellular retinoic acid-binding protein 2 (CRABP2), which delivers RA to the nuclear receptor RAR and whose mobilization to the nucleus is mediated by a 'classical-like' nuclear localization signal (NLS)...
September 8, 2016: Journal of Cell Science
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