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The Amyloid Precursor Protein is a conserved Wnt receptor bioRxiv. Cell Biol. Pub Date : 2021-01-18 Tengyuan Liu; Maya Nicolas; Tingting Zhang; Heather Rice; Alessia Soldano; Annelies Claeys; Iveta Petrova; Lee Fradkin; Bart De Strooper; Bassem A Hassan
The Amyloid Precursor Protein (APP) and its homologues are transmembrane proteins required for various aspects of neuronal development and activity, whose molecular function is unknown. Specifically, it is unclear whether APP acts as a receptor, and if so what its ligand(s) may be. We show that APP binds the Wnt ligands Wnt3a and Wnt5a and that this binding regulates APP protein levels. Wnt3a binding
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A non-olfactory shark adenosine receptor activates CFTR with unique pharmacology and structural features bioRxiv. Cell Biol. Pub Date : 2021-01-18 Sumeet Bhanot; Gabriele Hemminger; Cole L. Martin; Stephen G. Aller; John N. Forrest
Adenosine receptors (ADORs) are G-protein coupled purinoceptors that have several functions including regulation of chloride secretion via CFTR in human airway and kidney. We cloned an ADOR from Squalus acanthias (shark) that likely regulates CFTR in the rectal gland. Phylogenic- and expression- analyses indicate that elasmobranch ADORs are non-olfactory, and appear to represent extant predecessors
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Plasmodium falciparum Atg18 localizes to the food vacuole via interaction with the multi-drug resistance protein 1 and phosphatidylinositol 3-phosphate bioRxiv. Cell Biol. Pub Date : 2021-01-17 Renu Sudhakar; Divya Das; Subramanian Thanumalayan; Somesh Gorde; Puran Singh Sijwali
Autophagy is a lysosome-dependent degradative process involving over 35 Atg proteins. The autophagy repertoire in malaria parasites is limited and does not appear to be a major degradative process. To better understand the autophagy process, we investigated Plasmodium falciparum Atg18 (PfAtg18), a PROPPIN family protein, whose members like S. cerevisiae Atg18 (ScAtg18) and human WIPI2 are essential
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Regulation of neurofilament length and transport by a dynamic cycle of polymer severing and annealing bioRxiv. Cell Biol. Pub Date : 2021-01-17 Atsuko Uchida; Anthony Brown
We used long-term multi-field time-lapse imaging to analyze the movement, severing and annealing of single neurofilaments along axons of cultured neurons. All filaments were capable of rapid movement. However, long filaments paused and reversed more often, resulting in little net movement, whereas short filaments moved persistently for long distances, pausing and reversing less often. Long filaments
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Impaired mRNA splicing and proteostasis in preadipocytes in obesity-related metabolic disease bioRxiv. Cell Biol. Pub Date : 2021-01-17 Julia Sánchez-Ceinos; Rocío Guzmán-Ruiz; Oriol Alberto Rangel-Zuñiga; Elena Moreno-Caño; Mercedes del Río-Moreno; Elsa Maymo-Masip; Joan Vendrell; Sonia Fernández-Veledo; Jurga Laurencikiene; Mikael Rydén; Antonio Membrives; Raul M Luque; José López-Miranda; María M. Malagón
Preadipocytes are crucial for healthy adipose tissue expansion. Preadipocyte differentiation is altered in obese individuals, which has been proposed to contribute to obesity-associated metabolic disturbances. Here, we demonstrate that impaired alternative splicing and dysregulated endoplasmic reticulum (ER)-associated protein degradation (ERAD) represent marker pathways of dysfunctional preadipocytes
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SCFFbxw5 targets MCAK in G2/M to facilitate ciliogenesis in the following cell cycle bioRxiv. Cell Biol. Pub Date : 2021-01-17 Jörg Schweiggert; Gregor Habeck; Sandra Hess; Felix Mikes; Klaus Meese; Shoji Hata; Klaus-Peter Knobeloch; Frauke Melchior
The microtubule depolymerase Kif2C/MCAK plays important roles in various cellular processes and is frequently overexpressed in different cancer types. Despite the importance of its correct abundance, remarkably little is known about how MCAK levels are regulated in cells. Using comprehensive screening on protein microarrays, we identified 161 candidate substrates of the multi-subunit ubiquitin E3 ligase
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Cevipabulin-tubulin complex reveals a novel agent binding site on α-tubulin with tubulin degradation effect bioRxiv. Cell Biol. Pub Date : 2021-01-17 Jianhong Yang; Yamei Yu; Yong Li; Wei Yan; Haoyu Ye; Lu Niu; Minghai Tang; Zhoufeng Wang; Zhuang Yang; Heying Pei; Haoche Wei; Min Zhao; Jiaolin Wen; Linyu Yang; Liang Ouyang; Yuquan Wei; Qiang Chen; Weimin Li; Lijuan Chen
Microtubule, composed of αβ-tubulin heterodimers, remains as one of the most popular anticancer targets for decades. To date, anti-microtubule drugs are mainly functionally divided into microtubule-destabilizing and microtubule-stabilizing agents while microtubule- or tubulin-degradation agents are rarely reported. Six known binding sites on tubulin dimer are identified with five sites on β-tubulin
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Endothelial NRP2 influences angiogenesis by regulating actin pattern development and α5-integrin-p-FAK complex recruitment to assembling adhesion sites bioRxiv. Cell Biol. Pub Date : 2021-01-17 Christopher J Benwell; James AGE Taylor; Stephen D Robinson
The ability to form a variety of cell-matrix connections is crucial for angiogenesis to take place. Without stable anchorage to the extracellular matrix (ECM), endothelial cells (ECs) are unable to sense, integrate and disseminate growth factor stimulated responses that drive growth of a vascular bed. Neuropilin-2 (NRP2) is a widely expressed membrane-bound multifunctional non-tyrosine kinase receptor
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Actin dependent membrane polarization reveals the mechanical nature of the neuroblast polarity cycle bioRxiv. Cell Biol. Pub Date : 2021-01-17 Bryce LaFoya; Ken Prehoda
The Par complex directs fate determinant segregation from the apical membrane of asymmetrically dividing Drosophila neuroblasts. While the physical interactions that recruit the Par complex have been extensively studied, little is known about how the membrane itself behaves during polarization. We examined the membrane dynamics of neuroblasts and surrounding cells with super-resolution imaging, revealing
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Salt Inducible Kinase activation and IRE1-dependent intracellular ATP depletion to form Sec bodies in Drosophila cells. bioRxiv. Cell Biol. Pub Date : 2021-01-17 Catherine Rabouille; Chujun Zhang; Wessel van Leeuwen; Marloes Blotenburg; Angelica Aguilera-Gomez; Sem Brussee; Harm H Kampinga
The phase separation of the non-membrane bound Sec bodies occurs in Drosophila S2 cells by coalescence of components of the ER exit sites under the stress of amino-acid starvation. Here we address which signaling pathways cause Sec body formation. We find that two pathways are critical. The first is a SIK dependent pathway induced by salt (NaCl) stress in a necessary and sufficient manner. The second
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Random sub-diffusion and capture of genes by the nuclear pore reduces dynamics and coordinates interchromosomal movement bioRxiv. Cell Biol. Pub Date : 2021-01-17 Michael Chas Sumner; Steven Torrisi; Donna G Brickner; Jason H Brickner
Hundreds of genes interact with the yeast nuclear pore complex (NPC), localizing at the nuclear periphery and clustering with co-regulated genes. Dynamic tracking of peripheral genes shows that they cycle on and off the NPC and that interaction with the NPC slows their sub-diffusive movement. Furthermore, NPC-dependent inter-chromosomal clustering leads to coordinated movement of pairs of loci separated
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Protein Phosphatase 1 regulates atypical chromosome segregation and cell polarity during mitotic and meiotic division in Plasmodium sexual stages bioRxiv. Cell Biol. Pub Date : 2021-01-17 Mohammad Zeeshan; Rajan Pandey; AMIT KUMAR SUBUDHI; David J.P. Ferguson; Gursimran Kaur; Ravish Rashpa; Raushan Nugmanova; Declan Brady; Andrew Bottrill; sue vaughan; Mathieu Brochet; Mathieu Bollen; Arnab Pain; Anthony A Holder; David S. Guttery; Rita Tewari
PP1 is a conserved eukaryotic serine/threonine phosphatase that regulates many aspects of mitosis and meiosis, often working in concert with other phosphatases, such as CDC14 and CDC25. The proliferative stages of the parasite life cycle include sexual development within the mosquito vector, with male gamete formation characterized by an atypical rapid mitosis, consisting of three rounds of DNA synthesis
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Proprotein Convertase Subtilisin/kexin Type 9 Links Inflammation to Vascular Endothelial Cell Dysfunction bioRxiv. Cell Biol. Pub Date : 2021-01-17 Thorsten M Leucker; Nuria Amat-Codina; Stephen Chelko; Gary Gerstenblith
Vascular endothelial cell (EC) dysfunction is a pathological mediator of he development, progression, and clinical manifestations of atherosclerotic disease. Inflammation is associated with EC dysfunction, but the responsible mechanisms are not well characterized. There is substantial evidence that serum proprotein convertase subtilisin/kexin type 9 (PCSK9) is increased in pro-inflammatory states and
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CYRI-A regulates macropinocytic cup maturation and mediates integrin uptake, limiting invasive migration bioRxiv. Cell Biol. Pub Date : 2021-01-16 Anh Hoang Le; Tamas Yelland; Nikki Paul; Loic Fort; Savvas Nikolaou; Shehab Ismail; Laura Machesky
The Scar/WAVE complex is the major driver of actin nucleation at the plasma membrane, resulting in lamellipodia and membrane ruffles. While lamellipodia aid migration, membrane ruffles can generate macropinosomes - cup-like structures - important for nutrient uptake and regulation of cell surface receptor levels. How macropinosomes are formed and the role of the actin machinery in their formation and
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Motor Protein MYO1C is Critical for Photoreceptor Opsin Trafficking and Visual Function bioRxiv. Cell Biol. Pub Date : 2021-01-15 Ashish Solanki; Stephen Walterhouse; Rene Martin; Elisabeth Obert; Ehtesham Arif; Bushra Rahman; Barbel Rohrer; Joshua H Lipschutz; Rupak D Mukherjee; Russell A Norris; Jeffery Sundstrom; Hans-Joachim Knolker; Shahid Husain; Manas R Biswal; Deepak Nihalani; Glenn P Lobo
Unconventional myosins linked to deafness are also proposed to play a role in retinal cell physiology. However, their direct role in photoreceptor function remains unclear. We demonstrate that systemic loss of the unconventional myosin MYO1C in mice specifically affected opsin trafficking, leading to loss of visual function. Electroretinogram analysis of Myo1c knockout (Myo1c-KO) mice showed a progressive
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High-throughput phenotypic screen for genetic modifiers in patient-derived OPA1 mutant fibroblasts identifies PGS1 as a functional suppressor of mitochondrial fragmentation bioRxiv. Cell Biol. Pub Date : 2021-01-15 Emma Cretin; Priscilla Lopes; Elodie Vimont; Takashi Tatsuta; Thomas Langer; Patrick Yu-Wai-Man; Pascal Reynier; Timothy Wai
Mutations affecting the mitochondrial fusion protein Optic Atrophy 1 (OPA1) cause autosomal dominant optic atrophy (DOA) – one of the most common form of mitochondrial disease. The majority of patients develop isolated optic atrophy, but about 20% of OPA1 mutation carriers manifest more severe neurological deficits as part of a “DOA+” phenotype. OPA1 deficiency causes mitochondrial fragmentation and
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Patrolling human SLE haematopoietic progenitors demonstrate enhanced extramedullary colonisation; implications for peripheral tissue injury bioRxiv. Cell Biol. Pub Date : 2021-01-15 Ioannis Kokkinopoulos; Aggelos Banos; Maria Grigoriou; Anastasia Filia; Theodora Manolakou; Thenis Alissafi; Nikolaos Malissovas; Ioannis Mitroulis; Panayotis Verginis; Dimitrios T Boumpas
Systemic Lupus erythematosus (SLE) is an autoimmune disease where bone-marrow-derived haematopoietic cells have a key role in its pathogenesis with accumulating evidence suggesting an aberrant function of haematopoietic stem/progenitor cells (HSPCs). By employing next-generation sequencing, we compared the gene transcription signatures of CD34+ HSPCs deriving from either the bone marrow or HSPCs patrolling
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Mitotic R-loops direct Aurora B kinase to maintain centromeric cohesion bioRxiv. Cell Biol. Pub Date : 2021-01-15 Erin C. Moran; Limin Liu; Ewelina Zasadzinska; Courtney A. Kestner; Ali Sarkeshik; Henry DeHoyos; John R. Yates; Daniel Foltz; P. Todd Stukenberg
Recent work has shown that R-loops exist at mitotic centromeres, but the function of these R-loops is not well understood. Here, we report that mitotic R-loops arise in distinct locations from those formed during interphase. They accumulate on chromosome arms in prophase, where they are quickly resolved and continue to be produced at repetitive sequences including centromeres during a mitotic stall
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Identification of Paired-related Homeobox Protein 1 as a key mesenchymal transcription factor in Idiopathic Pulmonary Fibrosis bioRxiv. Cell Biol. Pub Date : 2021-01-15 E. Marchal-Duval; M. Homps-Legrand; A. Froidure; M. Jaillet; M. Ghanem; L. Deneuville; A. Justet; A. Maurac; A. Vadel; E. Fortas; A. Cazes; A. Joannes; L. Giersch; H. Mal; P. Mordant; C.M. Mounier; K. Schirduan; M. Korfei; A. Gunther; B. Mari; F. Jaschinski; B. Crestani; A.A. Mailleux
Matrix remodeling is a salient feature of idiopathic pulmonary fibrosis (IPF). Targeting cells driving matrix production and remodeling could be a promising avenue for IPF treatment. Analysis of public transcriptomic database identified paired-related homeobox protein-1 (PRRX1) as an upregulated mesenchymal transcription factor (TF) in IPF. We confirmed that PRRX1 isoforms were upregulated in IPF lung
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Establishment and characterization of mammary organoids from non-traditional model organisms bioRxiv. Cell Biol. Pub Date : 2021-01-15 Arianna P. Bartlett; Gerlinde R. Van de Walle
Mammary organoid (MaO) models are only available for a few traditional model organisms, limiting our ability to investigate mammary gland development and cancer across the diverse taxa of mammals. For example, horses are mammals with a similar mammary anatomy and function as humans, but they have a remarkably low incidence of mammary cancer, making the development of MaOs in non-traditional model organisms
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Pulsatile actomyosin contractions underlie Par polarity during the neuroblast polarity cycle bioRxiv. Cell Biol. Pub Date : 2021-01-15 Chet Huan Oon; Kenneth E. Prehoda
The Par complex is polarized to the apical cortex of asymmetrically dividing Drosophila neuroblasts. Previously we showed that Par proteins are polarized by apically directed cortical movements that require F-actin (Oon and Prehoda, 2019). Here we report the discovery of cortical actin pulses that begin before the Par complex is recruited to the cell cortex and ultimately become tightly coupled to
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Muscle-specific Cavin4 interacts with Bin1 to promote T-tubule formation and stability in developing skeletal muscle bioRxiv. Cell Biol. Pub Date : 2021-01-15 Harriet P. Lo; Ye-Wheen Lim; Zherui Xiong; Nick Martel; Charles Ferguson; Nicholas R. Ariotti; Jean Giacomotto; James A. Rae; Matthias Floetenmeyer; Shayli Varasteh Moradi; Ya Gao; Vikas A. Tillu; Di Xia; Huang Wang; Samira Rahnama; Susan J. Nixon; Michele Bastiani; Ryan D. Day; Kelly A. Smith; Nathan J. Palpant; Wayne A. Johnston; Kirill Alexandrov; Brett M. Collins; Thomas E. Hall; Robert G. Parton
The cavin proteins are essential for caveola biogenesis and function. Here, we identify a role for the muscle-specific component, Cavin4, in skeletal muscle T-tubule development by analyzing two vertebrate systems: mouse and zebrafish. In both models Cavin4 localized to T-tubules and loss of Cavin4 resulted in aberrant T-tubule maturation. In zebrafish, which possess duplicated cavin4 paralogs, Cavin4b
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Adiponectin Preserves Metabolic Fitness During Aging bioRxiv. Cell Biol. Pub Date : 2021-01-15 Na Li; Zhuzhen Zhang; Shangang Zhao; Yi Zhu; Christy M. Gliniak; Lavanya Vishvanath; Yu A. An; May-yun Wang; Yingfeng Deng; Qingzhang Zhu; Toshiharu Onodera; Orhan K Oz; Ruth Gordillo; Rana K. Gupta; Ming Liu; Tamas L. Horvath; Vishwa Deep Dixit; Philipp E. Scherer
Adiponectin is essential for the regulation of tissue substrate utilization and systemic insulin sensitivity. Clinical studies have suggested a positive association of circulating adiponectin with healthspan and lifespan. However, the direct effects of adiponectin on promoting healthspan and lifespan remain unexplored. Here, we are using an adiponectin null mouse and a transgenic adiponectin overexpression
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Mitochondrial Aurora kinase A induces mitophagy by interacting with MAP1LC3 and Prohibitin 2 bioRxiv. Cell Biol. Pub Date : 2021-01-15 Giulia Bertolin; Marie-Clotilde Alves-Guerra; Angelique Cheron; Agnes Burel; Claude Prigent; Roland Le Borgne; Marc Tramier
Epithelial and haematologic tumours often show the overexpression of the serine/threonine kinase AURKA. Recently, AURKA was shown to localise at mitochondria, where it regulates mitochondrial dynamics and ATP production. Here we define the molecular mechanisms of AURKA in regulating mitochondrial turnover by mitophagy. When overexpressed, AURKA induces the disappearance of Outer Mitochondrial Membrane
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Molecular mechanisms underlying attenuation of live attenuated Japanese encephalitis virus vaccine SA14-14-2 bioRxiv. Cell Biol. Pub Date : 2021-01-14 Pooja Hoovina Venkatesh; Saurabh Kumar; Naveen Kumar; Krishna Chaitanya; Lance Turtle; Vijaya Satchidanandam
The live attenuated Japanese encephalitis virus vaccine SA14-14-2 demonstrated ≥ 95% efficacy and is today the vaccine of choice against JEV globally. Relative to its parent strain SA14, SA14-14-2 carries 46 nucleotide and 24 amino acid alterations, with 8 of the latter located within the envelope glycoprotein. The vaccine strain also fails to synthesize the nonstructural protein NS1’ owing to a silent
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Dihydroceramide desaturase directs the switch between exosome production and autophagy for neuronal maintenance bioRxiv. Cell Biol. Pub Date : 2021-01-14 Chen-Yi Wu; Jhih-Gang Jhang; Chih-Wei Lin; Han-Chen Ho; Chih-Chiang Chan; Shu-Yi Huang
Exosomes play important roles in the nervous system. Mutations in the human dihydroceramide desaturase gene, DEGS1, are recently linked to severe neurological disorders, but the cause remains unknown. Here, we show that Ifc is required for the morphology and function of Drosophila photoreceptor neurons and not in the surrounding glia, but the degeneration of ifc-KO eyes can be rescued by glial expression
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The TPLATE subunit is essential for structural assembly of the endocytic TSET complex bioRxiv. Cell Biol. Pub Date : 2021-01-14 Klaas Yperman; Jie Wang; Dominique Eeckhout; Joanna Winkler; Lam Vu; Michael Vandorpe; Peter Grones; Evelien Mylle; Michael Kraus; Romain Merceron; Jonah Nolf; Eliana Mor; Pieter De Bruyn; Remy Loris; Martin Potocky; Savvas N. Savvides; Bert De Rybel; Geert De Jaeger; Daniel Van Damme; Roman Pleskot
All eukaryotic cells rely on endocytosis to regulate the plasma membrane proteome and lipidome. Most eukaryotic groups, with the exception of fungi and animals, have retained the evolutionary ancient TSET complex as a regulator of endocytosis. Despite the presence of similar building blocks in TSET, compared to other coatomer complexes, structural insight into this adaptor complex is lacking. Here
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The Necroptosis Effector MLKL drives Small Extracellular Vesicle Release and Tumour Growth in Glioblastoma bioRxiv. Cell Biol. Pub Date : 2021-01-14 Gwennan Andre-Gregoire; Tiphaine Douanne; An Thys; Clement Maghe; Kathryn A Jacobs; Cyndie Ballu; Kilian Trillet; Ignacio Busnelli; Vincent Hyenne; Jacky G. GOETZ; Nicolas Bidere; Julie Gavard
Extracellular vesicles (EVs) are lipid-based nano-sized particles that convey biological material from donor to recipient cells. They play key roles in tumour progression, notably in glioblastoma in which the subpopulation of Glioblastoma Stem-like Cells (GSCs) might represent a meaningful source of tumour-derived EVs. However, the mechanisms involved in the production and release of EVs by GSCs are
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Constitutive high expression of NOXA sensitizes human embryonic stem cells for rapid cell death bioRxiv. Cell Biol. Pub Date : 2021-01-14 Richa Basundra; Sahil Kapoor; Emilie Hollville; Nazanin Kiapour; Adriana Beltran Lopez; Nicole Marie Melchiorre; Mohanish Deshmukh
Human embryonic stem (hES) cells are highly sensitive to apoptotic stimuli such as DNA damage, which allows for rapid elimination of mutated cells during development. However, the mechanisms that maintain hES cells in the primed apoptotic state are not completely known. Key activators of apoptosis, the BH3-only proteins, are present at low levels in most cell types. In contrast, hES cells have constitutive
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Effects of arsenic on the topology and solubility of promyelocytic leukemia (PML)-nuclear bodies bioRxiv. Cell Biol. Pub Date : 2021-01-14 Seishiro Hirano; Osamu Udagawa
Promyelocytic leukemia (PML) proteins are involved in the pathogenesis of acute promyelocytic leukemia (APL). Trivalent arsenic (As3+) is known to cure APL by binding to cysteine residues of PML and enhance the degradation of PML-retinoic acid receptor (RAR), a t(15;17) gene translocation product in APL cells, and restore PML-nuclear bodies (NBs). The size, number, and shape of PML-NBs vary among cell
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FOXO1 and FOXO3 cooperatively regulate innate lymphoid cell development bioRxiv. Cell Biol. Pub Date : 2021-01-14 Thuy Luu Thanh; Jonas Norskov; Lucia Pena Perez; Shabnam Kharazi; Aleksandra Krstic; Stephan Meinke; Laurent Schmied; Nicolai Frengen; Yaser Heshmati; Marcin Kierczak; Thibault Bouderlique; Arnika Wagner; Charlotte Gustafsson; Benedict Chambers; Adnane Achour; Claudia Kutter; Petter Hoglund; Robert Mansson; Nadir Kadri
The natural killer (NK) and non-cytotoxic innate lymphoid cells (ILC) lineages play vital role in the regulation of the immune system. Yet understanding of mechanisms controlling NK/ILC development remains incomplete. The evolutionary conserved FOXO family of forkhead transcription factors are critical regulators of cellular processes. We found that the loss of FOXO1 and FOXO3 together caused impaired
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The Amot/Integrin protein complex transmits mechanical forces required for vascular expansion bioRxiv. Cell Biol. Pub Date : 2021-01-14 Yuanyuan Zhang; Yumeng Zhang; Sumako Kameishi; Giuseppina Barutello; Yujuan Zheng; Nicholas P. Tobin; John Nicosia; Katharina Hennig; David Kung-Chun Chiu; Martial Balland; Thomas H. Barker; Federica Cavallo; Lars Holmgren
Vascular development is a complex multistep process involving the coordination of cellular functions such as migration, proliferation and differentiation. Understanding the underlying mechanisms of these processes is of importance due to involvement of vessel expansion in various pathologies. How mechanical forces generated by cells and transmission of these physical forces control vascular development
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α-actinin-4 drives invasiveness by regulating myosin IIB expression and myosin IIA localization bioRxiv. Cell Biol. Pub Date : 2021-01-12 Amlan Barai; Abhishek Mukherjee; Alakesh Das; Neha Saxena; Shamik Sen
The mechanisms by which the mechanoresponsive actin crosslinking protein α-actinin-4 (ACTN4) regulates cell motility and invasiveness remains incompletely understood. Here we show that in addition to regulating protrusion dynamics and focal adhesion formation, ACTN4 transcriptionally regulates expression of non-muscle myosin IIB (NMM IIB), which is essential for mediating nuclear translocation during
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Enrichment of Skeletal Stem Cells from Human Bone Marrow Using Spherical Nucleic Acids bioRxiv. Cell Biol. Pub Date : 2021-01-14 Miguel Xavier; Marilena Kyriazi; Stuart Lanham; Konstantina Alexaki; Afaf El-Sagheer; Tom Brown; Antonios Kanaras; Richard O. C. Oreffo
Human bone marrow (BM) derived stromal cells contain a population of skeletal stem cells (SSCs), with the capacity to differentiate along the osteogenic, adipogenic and chondrogenic lineages enabling their application to clinical therapies. However, current methods, to isolate and enrich SSCs from human tissues remain, at best, challenging in the absence of a specific SSC marker. Unfortunately, none
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Standardized quality control workflow to evaluate the reproducibility and differentiation potential of human iPSCs into neurons bioRxiv. Cell Biol. Pub Date : 2021-01-14 Carol X.-Q. Chen; Narges Abdian; Gilles Maussion; Rhalena A. Thomas; Iveta Demirova; Eddie Cai; Mahdieh Tabatabaei; Lenore K. Beitel; Jason Karamchandani; Edward A. Fon; Thomas M Durcan
Induced pluripotent stem cells (iPSCs) derived from human somatic cells have created new opportunities to generate disease-relevant cells. Thus, as the use of patient-derived stem cells has become more widespread, having a workflow to monitor each line is critical. This ensures iPSCs pass a suite of quality control measures, promoting reproducibility across experiments and between labs. With this in
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Real-time luminescence assay for cytoplasmic cargo delivery of extracellular vesicles bioRxiv. Cell Biol. Pub Date : 2021-01-13 Masaharu Somiya; Shun'ichi Kuroda
Extracellular vesicles (EVs) have been considered to deliver biological cargos between cells and mediate intercellular communication. However, the mechanisms that underlie the biological process of EV uptake and cytoplasmic cargo release in recipient cells are largely unknown. Quantitative and real-time assays for assessment of the cargo delivery efficiency inside recipient cells have not been feasible
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A role for SHARPIN in platelet linear protein ubiquitination and function bioRxiv. Cell Biol. Pub Date : 2021-01-13 Samantha F Moore; Xiaojuan Zhao; Shahida Mallah; Alastair W Poole; Stuart J Mundell; James L Hutchinson; Ingeborg Hers
SHARPIN (Src homology 3 and multiple ankyrin repeat domains protein (SHANK)-associated RH domain-interacting protein) as part of the linear ubiquitin chain assembly complex (LUBAC) catalyses the addition of linear (Met1-linked) ubiquitin chains to substrates. As part of this complex SHARPIN acts as a multi-functional modulator of immune/inflammatory responses through regulation of NfkB activation.
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Periplasm homeostatic regulation maintains spatial constraints essential for cell envelope processes and cell viability bioRxiv. Cell Biol. Pub Date : 2021-01-13 Eric Mandela; Christopher J Stubenrauch; David Ryoo; Hyea Hwang; Eli J Cohen; Von Vergel L Torres; Pankaj Deo; Chaille T Webb; Cheng Huang; Ralf B Schittenhelm; Morgan Beeby; J. C. Gumbart; Trevor Lithgow; Iain D Hay
The cell envelope of Gram-negative bacteria consists of two membranes surrounding a periplasm and peptidoglycan layer. Molecular machines spanning the cell envelope depend on spatial constraints and load-bearing forces across the cell envelope and surface. The mechanisms dictating spatial constraints across the cell envelope remain incompletely defined. In Escherichia coli, the coiled-coil lipoprotein
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Dynlt3, a novel fundamental regulator of melanosome movement, distribution, maturation and transfer bioRxiv. Cell Biol. Pub Date : 2021-01-13 Zackie Aktary; Alejandro Conde; Florian Rambow; Mathilde Di Marco; Francois Amblard; Ilse Hurbain; Graca Raposo; Cedric Delevoye; Sylvie Coscoy; Lionel LARUE
Skin pigmentation is dependent on cellular processes including melanosome biogenesis, transport, maturation and transfer to keratinocytes. However, how the cells finely control these processes in space and time to ensure proper pigmentation remains unclear. Here, we show that a component of the cytoplasmic dynein complex, Dynlt3, is required for efficient melanosome transport, maturation and transfer
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Microtubule associated proteins and motors required for ectopic microtubule array formation in S. cerevisiae bioRxiv. Cell Biol. Pub Date : 2021-01-13 Brianna R King; Janet B Meehl; Tamira Vojnar; Mark Winey; Eric G Muller; Trisha N Davis
The mitotic spindle is resilient to perturbation due to the concerted, and sometimes redundant, action of motors and microtubule-associated proteins. Here we utilize an inducible ectopic microtubule nucleation site in the nucleus of Saccharomyces cerevisiae to study three necessary steps in the formation of a bipolar array: the recruitment of the gamma-tubulin complex, nucleation and elongation of
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Robust integrated intracellular organization of the human iPS cell: where, how much, and how variable bioRxiv. Cell Biol. Pub Date : 2021-01-13 Matheus P. Viana; Jianxu Chen; Theo A. Knijnenburg; Ritvik Vasan; Calysta Yan; Joy E. Arakaki; Matte Bailey; Ben Berry; Antoine Borensztejn; Jackson M. Brown; Sara Carlson; Julie A. Cass; Basudev Chaudhuri; Kimberly R. Cordes Metzler; Mackenzie E. Coston; Zach J. Crabtree; Steve Davidson; Colette M. DeLizo; Shailja Dhaka; Stephanie Q. Dinh; Thao P. Do; Justin Domingus; Rory M. Donovan-Maiye; Tyler
Despite the intimate link between cell organization and function, the principles underlying intracellular organization and the relation between organization, gene expression and phenotype are not well understood. We address this by creating a benchmark for mean cell organization and the natural range of cell-to-cell variation. This benchmark can be used for comparison to other normal or abnormal cell
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Visualizing protein-protein interactions in plants by rapamycin-dependent delocalization bioRxiv. Cell Biol. Pub Date : 2021-01-13 Joanna Winkler; Evelien Mylle; Andreas De Meyer; Benjamin Pavie; Julie Merchie; Peter Grones; Daniel Van Damme
Identifying protein-protein interactions (PPI) is crucial for understanding biological processes. Many PPI tools are available, yet only some function within the context of a plant cell. Narrowing down even further, only a few tools allow complex multiprotein interactions to be visualized. Here, we present a conditional in vivo PPI tool for plant research that meets these criteria. Knocksideways in
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ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells bioRxiv. Cell Biol. Pub Date : 2021-01-12 Maria F Ullo; Jeremy S Logue
Melanoma cells have been shown to undergo fast amoeboid or Leader Bleb-Based Migration (LBBM), requiring a large and stable bleb for migration. In leader blebs, is a rapid flow of cortical actin that drives the cell forward. Here, we tested the hypothesis that the actin severing factors, ADF and cofilin-1, are essential for contractility and actin flow for LBBM. Using RNAi in melanoma A375 cells, we
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Restore mitophagy is essential to prevent cardiac oxidative stress during hypertrophy bioRxiv. Cell Biol. Pub Date : 2021-01-12 Victoriane Peugnet; Maggy Chwastyniak; Steve Lancel; Laurent Bultot; Natacha Fourny; Olivia Beseme; Anne Loyens; Wilfried Heyse; Philippe Amouyel; Luc Bertrand; Florence Pinet; Emilie Dubois-Deruy
Heart failure, mostly associated with cardiac hypertrophy, is still a major cause of illness and death. Oxidative stress causes contractile failure and the accumulation of reactive oxygen species leads to mitochondrial dysfunction, associated with aging and heart failure, suggesting that mitochondria-targeted therapies could be effective in this context. The purpose of this work was to characterize
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SLO2.1 and NALCN form a functional complex to modulate myometrial cell excitability bioRxiv. Cell Biol. Pub Date : 2021-01-12 Juan J Ferreira; Chinwendu Amazu; Lis C Puga-Molina; Xiaofeng Ma; Sarah K England; Celia M Santi
Depolarization of the myometrial smooth muscle cell (MSMC) resting membrane potential is necessary for the transition of the uterus from a quiescent state to a contractile state. The molecular mechanisms involved in this transition are not completely understood. Here, we report a novel coupled system between the Na+-activated K+ channel (SLO2.1) and the non-selective Na+ leak channel (NALCN) which
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Myofibrillar Structural Variability Underlies Contractile Function in Stem Cell-Derived Cardiomyocytes bioRxiv. Cell Biol. Pub Date : 2021-01-12 Kathryn Ufford; Sabrina Friedline; Zhaowen Tong; Vi T Tang; Amani S Dobbs; Yao-Chang Tsan; Stephanie L Bielas; Allen P Liu; Adam S Helms
Disease modeling and pharmaceutical testing using cardiomyocytes derived from induced pluripotent stem cell (iPSC-CMs) requires accurate assessment of contractile function. Micropatterning iPSC-CMs on elastic substrates controls cell shape and alignment to enable contractile studies, but determinants of intrinsic variability in this system have been incompletely characterized. The objective of this
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Restored TDCA and Valine Levels Imitate the Effects of Bariatric Surgery bioRxiv. Cell Biol. Pub Date : 2021-01-12 Stefan G. Tullius; Markus Quante; Jasper Iske; Timm Heinbokel; Bhavna N. Desai; Hector Rodriguez Cetina Biefer; Yeqi Nian; Felix Krenzien; Hirofumi Uehara; Ryoichi Maenosono; Haruhito Azuma; Johann Pratschke; Christine Falk; Tammy Lo; Eric Sheu; Ali Tavakkoli; David Perkins; Maria-Luisa Alegre; Alexander S. Banks; Abdallah Elkhal
Obesity is widespread and linked to various co-morbidities. Bariatric surgery has been identified as the only effective treatment, promoting sustained weight loss and the remission of co-morbidities. Mass-spectrometric metabolomic profiling revealed significantly reduced systemic levels of TDCA and L-valine in Diet-Induced-Obese (DIO) mice. TDCA and L-Valine levels were restored after sleeve gastrectomies
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Dissecting lipid contents in the distinct regions of native retinal rod disk membranes bioRxiv. Cell Biol. Pub Date : 2021-01-11 Christopher L. Sander; Avery E. Sears; Antonio F. M. Pinto; Elliot H. Choi; Shirin L. Kahremany; Hui Grace Jin; Susie Suh; Els Pardon; Zhiqian Dong; Jan Steyaert; Alan Saghatelian; Dorota Skowronska-Krawczyk; Philip D. Kiser; Krzysztof Palczewski
Photoreceptors rely on distinct membrane compartments to support their specialized function. Unlike protein localization, identification of critical differences in membrane content has not yet been expanded to lipids, due to the difficulty of isolating domain-specific samples. We have overcome this by using SMA to co-immunopurify membrane proteins and their native lipids from two regions of photoreceptor
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Forkhead box O1-mediated ubiquitination suppresses RIG-I-mediated antiviral immune responses via its DNA binding domain bioRxiv. Cell Biol. Pub Date : 2021-01-11 Wei Liu; Yanlei Yue; Wenwen Zhang; Zhenling Ma; Wen Yao; Tao Li; Jun Xu; Wenqing Li; Lei Sun; Wenjun Liu; Guoqing Yang
The authors have withdrawn our manuscript whilst we perform additional experiments to test some of our conclusions further. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.
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Energy Expenditure during Cell Spreading Regulates the Stem Cells Responses to Matrix Stiffness bioRxiv. Cell Biol. Pub Date : 2021-01-11 Jing Xie; Min Bao; Xinyu Hu; Werner J. H. Koopman; Wilhelm T. S. Huck
Cells respond to the mechanical properties of the extracellular matrix (ECM) through the formation of focal adhesions (FAs), re-organization of the actin cytoskeleton, and adjustment of cell contractility. These are energy-demanding processes, but a potential causality between mechanical cues (matrix stiffness) and cellular (energy) metabolism remains largely unexplored. Here, we culture human mesenchymal
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Gene family evolution underlies cell type diversification in the hypothalamus of teleosts bioRxiv. Cell Biol. Pub Date : 2021-01-11 Maxwell E.R. Shafer; Ahilya N. Sawh; Alex F. Schier
Hundreds of cell types form the vertebrate brain, but it is largely unknown how similar these cellular repertoires are between or within species, or how cell type diversity evolves. To examine cell type diversity across and within species, we performed single-cell RNA sequencing of ~130,000 hypothalamic cells from zebrafish (Danio rerio) and surface- and cave-morphs of Mexican tetra (Astyanax mexicanus)
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Hypoxia reduces cell attachment of SARS-CoV-2 spike protein by modulating the expression of ACE2 and heparan sulfate bioRxiv. Cell Biol. Pub Date : 2021-01-11 Endika Prieto-Fernandez; Leire Egia-Mendikute; Laura Vila-Vecilla; So Young Lee; Alexandre Bosch; Adrian Barreira-Manrique; Ana Garcia del Rio; Asier Antonana-Vildosola; Borja Jimenez-Lasheras; Asis Palazon
A main clinical parameter of Covid-19 pathophysiology is hypoxia. Here we show that hypoxia decreases the attachment of the receptor binding domain (RBD) and the S1 subunit (S1) of the spike protein to epithelial cells. In Vero E6 cells, hypoxia reduces the protein levels of ACE2, which might in part explain the observed reduction of the infection rate. However, hypoxia also inhibits the binding of
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Free-fatty acid receptor 4 inhibitory signaling in delta cells regulates islet hormone secretion in mice bioRxiv. Cell Biol. Pub Date : 2021-01-11 Marine L Croze; Marcus Flisher; Arthur Guillaume; Caroline Tremblay; Glyn M Noguchi; Sabrina Granziera; Kevin Vivot; Vincent C Castillo; Scott A Campbell; Julien Ghislain; Mark O Huising; Vincent Poitout
Objective: Maintenance of glucose homeostasis requires the precise regulation of hormone secretion from the endocrine pancreas. Free fatty-acid receptor 4 (FFAR4/GPR120) is a G protein-coupled receptor whose activation in islets of Langerhans promotes insulin and glucagon secretion and inhibits somatostatin secretion. However, the contribution of individual islet cell types (alpha, beta, and delta
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Nuclear m6A reader Ythdc1 regulates the scaffold function of LINE1 in mouse ESCs bioRxiv. Cell Biol. Pub Date : 2021-01-11 Chuan Chen; Wenqiang Liu; Jiayin Guo; Yuanyuan Liu; Xuelian Liu; Jun Liu; Xiaoyang Dou; Rongrong Le; Yixin Huang; Chong Li; Lingyue Yang; Xiaochen Kou; Yanhong Zhao; You Wu; Jiayu Chen; Hong Wang; Bin Shen; Yawei Gao; Shaorong Gao
N6-methyladenosine (m6A) on chromosome-associated regulatory RNAs (carRNAs), including repeat RNAs, play important roles in tuning the chromatin state and transcription. Among diverse RNA-chromatin interacting modes, the nuclear RNA scaffold is considered important for trans-interactions but has not yet been connected with m6A yet. Here, we found that Ythdc1 played indispensable roles in the embryonic
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Mitofusins 1 and 2 collaborate to fuel pancreatic beta cell insulin release via regulation of both mitochondrial structure and DNA content bioRxiv. Cell Biol. Pub Date : 2021-01-11 Vaibhav Sidarala; Jie Zhu; Gemma L. Pearson; Emma C. Reck; Brett A Kaufman; Scott A Soleimanpour
Mitochondria are vital for β-cell function, yet the importance of mitochondrial fusion for glucose homeostasis is uncertain. Here, we report that the dynamin-like GTPases Mitofusin 1 and 2 (Mfn1 and Mfn2) are critical for glucose-stimulated insulin secretion (GSIS). Whereas Mfn1 and Mfn2 individually were dispensable for glucose control, combined Mfn1/2 deletion in β-cells induced mitochondrial fragmentation
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In-situ observation of mitochondrial biogenesis as the early event of apoptosis bioRxiv. Cell Biol. Pub Date : 2021-01-11 Chang-sheng Shao; Xiu-Hong Zhou; Yu-Hui Miao; Peng Wang; Qian-qian Zhang; Qing Huang
Mitochondrial biogenesis is a cell response to external stimuli which is generally believed to suppress apoptosis. However, during the process of apoptosis, whether mitochondrial biogenesis occurs in the early stage of the apoptotic cells remains unclear. To address this question, we constructed the COX8-EGFP-ACTIN-mCherry HeLa cells with recombinant fluorescent proteins respectively tagged on the
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Imaging cytoplasmic lipid droplets in vivo with fluorescent perilipin 2 and perilipin 3 knockin zebrafish bioRxiv. Cell Biol. Pub Date : 2021-01-10 Meredith H Wilson; Stephen C Ekker; Steven A. Farber
Cytoplasmic lipid droplets are highly dynamic storage organelles; their rapid synthesis, expansion, and degradation, as well as their varied interactions with other organelles allow cells to maintain lipid homeostasis. While the molecular details of lipid droplet dynamics are currently a very active area of investigation, this work has been primarily performed in cultured cells and in vitro systems
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Resolving the graft ischemia-reperfusion injury during liver transplantation at the single cell resolution bioRxiv. Cell Biol. Pub Date : 2021-01-10 Linhe Wang; Jie Li; Shuai He; Haitian Chen; Shujiao He; Meixian Yin; Dawei Zou; Jinghong Xu; Shirui Chen; Tao Luo; Xinyu Yu; JINXIN BEI; Zhiyong Guo; Xiaoshun He
Ischemia-reperfusion injury (IRI) remains the major reason for impaired donor graft function and increased mortality post-liver transplantation. The mechanism of IRI involves multiple pathophysiological processes and numerous types of cells. However, a systematic and comprehensive single-cell transcriptional profile of intrahepatic cells during liver transplantation is still unclear. We performed a
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Single-cell RNA-seq analysis reveals penaeid shrimp hemocyte subpopulations and cell differentiation process bioRxiv. Cell Biol. Pub Date : 2021-01-10 Keiichiro Koiwai; Takashi Koyama; Soichiro Tsuda; Atsushi Toyoda; Kiyoshi Kikuchi; Hiroaki Suzuki; Ryuji Kawano
Crustacean aquaculture is expected to be a major source of fishery commodities in the near future. An immune priming system of shrimp is crucial for a sustainable supply, as shrimp do not have an adaptive immune system; however, little is known about their immunity. Hemocytes are known as key agents of the crustacean immune system; nevertheless, we have yet to identify the different cell types, functions