DOCSLIB.ORG

The Membrane Phospholipid Binding Protein Annexin A2 Promotes

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Membrane Phospholipid Binding Protein Annexin A2 Promotes The Membrane Phospholipid Binding Protein Annexin A2 Promotes Phagocytosis and Nonlytic Exocytosis of Cryptococcus neoformans and Impacts Survival in Fungal This information is current as Infection of September 23, 2021. Sabriya Stukes, Carolina Coelho, Johanna Rivera, Anne E. Jedlicka, Katherine A. Hajjar and Arturo Casadevall J Immunol published online 1 July 2016 http://www.jimmunol.org/content/early/2016/07/01/jimmun Downloaded from ol.1501855 Supplementary http://www.jimmunol.org/content/suppl/2016/07/01/jimmunol.150185 http://www.jimmunol.org/ Material 5.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 23, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published July 1, 2016, doi:10.4049/jimmunol.1501855 The Journal of Immunology The Membrane Phospholipid Binding Protein Annexin A2 Promotes Phagocytosis and Nonlytic Exocytosis of Cryptococcus neoformans and Impacts Survival in Fungal Infection Sabriya Stukes,* Carolina Coelho,*,† Johanna Rivera,* Anne E. Jedlicka,† Katherine A. Hajjar,‡,x and Arturo Casadevall*,† Cryptococcus neoformans is a fungal pathogen with a unique intracellular pathogenic strategy that includes nonlytic exocytosis, a phenomenon whereby fungal cells are expunged from macrophages without lysing the host cell. The exact mechanism and specific proteins involved in this process have yet to be completely defined. Using murine macrophages deficient in the membrane Downloaded from phospholipid binding protein, annexin A2 (ANXA2), we observed a significant decrease in both phagocytosis of yeast cells and the frequency of nonlytic exocytosis. Cryptococcal cells isolated from Anxa2-deficient (Anxa22/2) bone marrow–derived macro- phages and lung parenchyma displayed significantly larger capsules than those isolated from wild-type macrophages and tissues. Concomitantly, we observed significant differences in the amount of reactive oxygen species produced between Anxa22/2 and Anxa2+/+ macrophages. Despite comparable fungal burden, Anxa22/2 mice died more rapidly than wild-type mice when infected 2/2 with C. neoformans, and Anxa2 mice exhibited enhanced inflammatory responses, suggesting that the reduced survival http://www.jimmunol.org/ reflected greater immune-mediated damage. Together, these findings suggest a role for ANXA2 in the control of cryptococcal infection, macrophage function, and fungal morphology. The Journal of Immunology, 2016, 197: 000–000. ryptococcus neoformans is an environmental fungus with and laccase, which deactivate microbicidal compounds; and 3) the a complex intracellular pathogenic strategy that is inti- ability of the fungus to damage the phagosomal membrane allowing C mately related to its capacity for virulence (1–3). Fungal efflux of phagosomal contents into the cytoplasm (1, 8, 10–13). infection results from the inhalation of aerosolized infectious However, perhaps the most unusual aspect of the intracellular strat- particles (4). Unlike other intracellular pathogens, C. neoformans egy of this fungal pathogen is its ability to escape the macrophage can survive and replicate inside an acidic phagosome that un- in a process known as nonlytic exocytosis (14, 15). by guest on September 23, 2021 dergoes normal maturation (5). The outcome of the interaction Nonlytic exocytosis has been described in vitro for different cell between this fungal pathogen and macrophages appears to be a types such as J774 murine macrophage-like cells, primary mouse and critical determinant of virulence (3, 6–9). human macrophages, Drosophila S2 cells, and amoebae (4, 16–19). The mechanisms responsible for fungal intracellular survival Nonlytic exocytosis was shown to occur in vivo and could provide a involve a combination of factors that include: 1) residence in a large mechanism by which yeast cells migrating from the lung in Trojan phagosome where lysosomal contents are diluted; 2) the presence of horse macrophages are released into the circulation to infect the powerful antioxidant mechanisms including a large polysaccharide brain by transcytosis of the blood–brain barrier (20, 21). This in- capsule, cell wall–associated melanin that can absorb lysosome- teraction appears to be a pathogen-driven phenomenon because it generated oxidants and enzymes, such as superoxide dismutase requires cryptococcal cell viability and is selectively confined to fungal cells when macrophages ingest both live yeast cells and *Department of Microbiology and Immunology, Albert Einstein College of Medi- FITC-labeled beads (15). The phagosome appears to play a crucial cine, Bronx, NY 10461; †Johns Hopkins Bloomberg School of Public Health, Balti- role during this process, because phagosome labeling experiments more, MD 21205; ‡Department of Pediatrics, Weill Cornell Medicine, New York, NY x suggest that during nonlytic exocytosis, the entire organelle is ex- 10065; and Department of Cell and Developmental Biology, Weill Cornell Medi- cine, New York, NY 10065 punged, allowing for the release of fungal cells into the extracellular ORCIDs: 0000-0002-7523-3031 (C.C.); 0000-0001-9332-4861 (J.R.); 0000-0003- environment (22). Nonlytic exocytosis is under the control of both 3977-4356 (K.A.H.); 0000-0002-9402-9167 (A.C.). host and fungal factors, including host actin (22), phagosomal pH Received for publication August 18, 2015. Accepted for publication June 2, 2016. (20), fungal SEC14 (23), cytokine stimulation (24), host autophagy This work was supported by National Institutes of Health Grants 5R01A1033774, (25), and the presence of a capsule on fungal cells (15). 5R37AI033142, and 5T32A107506, and Clinical and Translational Science Awards In this study, we investigated the role of the membrane binding Grants 1 ULI TR001073-01, 1 TLI 1 TR001072-01, and 1 KL2 TR001071 from the National Center for Advancing Translational Sciences. protein annexin A2 (ANXA2) during in vitro fungal interactions with macrophages and in cryptococcal pathogenesis. Annexins are a Address correspondence and reprint requests to Dr. Arturo Casadevall, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205. family of calcium binding proteins that function by bringing cellular E-mail address: [email protected] membranes in close contact with each other to promote fusion (26). The online version of this article contains supplemental material. They function in a wide variety of biological processes, some of Abbreviations used in this article: ANXA2, annexin A2; BMDM, bone marrow– which include membrane trafficking, phagocytosis, the endocytic derived macrophage; ROS, reactive oxygen species; SNAP, soluble N-ethylmaleimide pathway, and of interest to us, exocytosis of secretory vesicles (27– sensitive fusion attachment protein. 29). Specifically, we focused on ANXA2 because this protein me- Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 diates endosomal membrane–membrane fusion and plays a role in www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501855 2 ANNEXIN A2 DEPLETION IN MACROPHAGES IMPACTS FUNGAL INFECTION the docking mechanism needed for vesicles to adhere to cellular Casadevall laboratory for several years. Unless otherwise specified, yeast membranes. ANXA2 is also hypothesized to function in the cells were washed three times in sterile PBS, counted, and used at an E:T membrane fusion events leading to the release of vesicles in chro- ratio of 5:1 for all in vitro experiments unless otherwise noted. maffin cells, exocytosis of lamellar bodies in alveolar epithelial C57BL/6J and Anxa22/2 mouse strains cells, and regulated exocytosis of von Willebrand factor pack- 2/2 Homozygous breeding pairs of Anxa2 knockout (Anxa2 ) mice from a aged in Weibel-Palade bodies in bovine endothelial cells (30– C57BL/6J background were obtained from the Hajjar laboratory (New 32). In addition, ANXA2 mediates secretion of vesical-bound York, NY) (36) and were maintained at the Animal Institute of Albert macromolecular collagen VI from bronchial epithelial cells (33). Einstein College of Medicine. Male and female C57BL/6J (8–10 wk old) There is minimal information linking ANXA2 and C. neoformans, mice were obtained from the National Cancer Institute (Fredrick, MD). In all experiments, mice were used at 8–10 wk of age. Mice were housed in and what has been shown mostly focuses on the interaction of fungal sterile microisolator cages in a barrier environment and were maintained cells and endothelial cells. Vu et al. showed that ANXA2 and in a specific pathogen-free barrier facility in microisolator cages, fed irra- S100A10 genes were both upregulated upon contact and ingestion of diated rodent food, provided with autoclaved bedding, and routinely moni- fungal cells by brain endothelial cells. This upregulation translated tored
Load more

Recommended publications
  • Annexin A2 Flop-Out Mediates the Non-Vesicular Release of Damps/Alarmins from C6 Glioma Cells Induced by Serum-Free Conditions
    cells Article Annexin A2 Flop-Out Mediates the Non-Vesicular Release of DAMPs/Alarmins from C6 Glioma Cells Induced by Serum-Free Conditions Hayato Matsunaga 1,2,† , Sebok Kumar Halder 1,3,† and Hiroshi Ueda 1,4,* 1 Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8521, Japan; [email protected] (H.M.); [email protected] (S.K.H.) 2 Department of Medical Pharmacology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8523, Japan 3 San Diego Biomedical Research Institute, San Diego, CA 92121, USA 4 Department of Molecular Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan * Correspondence: [email protected]; Tel.: +81-75-753-4536 † These authors contributed equally to this work. Abstract: Prothymosin alpha (ProTα) and S100A13 are released from C6 glioma cells under serum- free conditions via membrane tethering mediated by Ca2+-dependent interactions between S100A13 and p40 synaptotagmin-1 (Syt-1), which is further associated with plasma membrane syntaxin-1 (Stx-1). The present study revealed that S100A13 interacted with annexin A2 (ANXA2) and this interaction was enhanced by Ca2+ and p40 Syt-1. Amlexanox (Amx) inhibited the association between S100A13 and ANXA2 in C6 glioma cells cultured under serum-free conditions in the in situ proximity ligation assay. In the absence of Amx, however, the serum-free stress results in a flop-out of ANXA2 Citation: Matsunaga, H.; Halder, through the membrane, without the extracellular release. The intracellular delivery of anti-ANXA2 S.K.; Ueda, H. Annexin A2 Flop-Out antibody blocked the serum-free stress-induced cellular loss of ProTα, S100A13, and Syt-1.
    [Show full text]
  • Annexin A1 Is a New Functional Linker Between Actin Filaments and Phagosomes During Phagocytosis
    578 Research Article Annexin A1 is a new functional linker between actin filaments and phagosomes during phagocytosis Devang M. Patel1, Syed Furquan Ahmad1, Dieter G. Weiss1, Volker Gerke2 and Sergei A. Kuznetsov1,* 1Institute of Biological Sciences, Cell Biology and Biosystems Technology, University of Rostock, Albert-Einstein Straße 3, Rostock 18059, Germany 2Institute of Medical Biochemistry, Centre for Molecular Biology of Inflammation, University of Münster, Von-Esmarch-Straße 56, Münster 48149, Germany *Author for correspondence ([email protected]) Accepted 19 October 2010 Journal of Cell Science 124, 578-588 © 2011. Published by The Company of Biologists Ltd doi:10.1242/jcs.076208 Summary Remodelling of the actin cytoskeleton plays a key role in particle internalisation and the phagosome maturation processes. Actin- binding proteins (ABPs) are the main players in actin remodelling but the precise role of these proteins in phagocytosis needs to be clarified. Annexins, a group of ABPs, are known to be present on phagosomes. Here, we identified annexin A1 as a factor that binds to isolated latex bead phagosomes (LBPs) in the presence of Ca2+ and facilitates the F-actin–LBP interaction in vitro. In macrophages the association of endogenous annexin A1 with LBP membranes was strongly correlated with the spatial and temporal accumulation of F-actin at the LBP. Annexin A1 was found on phagocytic cups and around early phagosomes, where the F-actin was prominently concentrated. After uptake was completed, annexin A1, along with F-actin, dissociated from the nascent LBP surface. At later stages of phagocytosis annexin A1 transiently concentrated only around those LBPs that showed transient F-actin accumulation (‘actin flashing’).
    [Show full text]
  • 2812 Matrix Vesicles: Structure, Composition, Formation and Function in Ca
    [Frontiers in Bioscience 16, 2812-2902, June 1, 2011] Matrix vesicles: structure, composition, formation and function in calcification Roy E. Wuthier Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208 TABLE OF CONTENTS 1. Abstract 2. Introduction 3. Morphology of matrix vesicles (MVs) 3.1. Conventional transmission electron microscopy 3.2. Cryofixation, freeze-substitution electron microscopy 3.3. Freeze-fracture studies 4. Isolation of MVs 4.1. Crude collagenase digestion methods 4.2. Non-collagenase dependent methods 4.3. Cell culture methods 4.4. Modified collagenase digestion methods 4.5. Other isolation methods 5. MV proteins 5.1. Early SDS-PAGE studies 5.2. Isolation and identification of major MV proteins 5.3. Sequential extraction, separation and characterization of major MV proteins 5.4. Proteomic characterization of MV proteins 6. MV-associated extracellular matrix proteins 6.1. Type VI collagen 6.2. Type X collagen 6.3. Proteoglycan link protein and aggrecan core protein 6.4. Fibrillin-1 and fibrillin-2 7. MV annexins – acidic phospholipid-dependent ca2+-binding proteins 7.1. Annexin A5 7.2. Annexin A6 7.3. Annexin A2 7.4. Annexin A1 7.5. Annexin A11 and Annexin A4 8. MV enzymes 8.1. Tissue-nonspecific alkaline phosphatase(TNAP) 8.1.1. Molecular structure 8.1.2. Amino acid sequence 8.1.3. 3-D structure 8.1.4. Disposition in the MV membrane 8.1.5. Catalytic properties 8.1.6. Collagen-binding properties 8.2. Nucleotide pyrophosphate phosphodiesterase (NPP1, PC1) 8.3. PHOSPHO-1 (Phosphoethanolamine/Phosphocholine phosphatase 8.4. Acid phosphatase 8.5.
    [Show full text]
  • Changes in the Cytosolic Proteome of Aedes Malpighian Tubules
    329 The Journal of Experimental Biology 212, 329-340 Published by The Company of Biologists 2009 doi:10.1242/jeb.024646 Research article Signaling to the apical membrane and to the paracellular pathway: changes in the cytosolic proteome of Aedes Malpighian tubules Klaus W. Beyenbach1,*, Sabine Baumgart2, Kenneth Lau1, Peter M. Piermarini1 and Sheng Zhang2 1Department of Biomedical Sciences, VRT 8004, Cornell University, Ithaca, NY 14853, USA and 2Proteomics and Mass Spectrometry Core Facility, 143 Biotechnology Building, Cornell University, Ithaca, NY 14853, USA *Author for correspondence (e-mail: [email protected]) Accepted 6 November 2008 Summary Using a proteomics approach, we examined the post-translational changes in cytosolic proteins when isolated Malpighian tubules of Aedes aegypti were stimulated for 1 min with the diuretic peptide aedeskinin-III (AK-III, 10–7 mol l–1). The cytosols of control (C) and aedeskinin-treated (T) tubules were extracted from several thousand Malpighian tubules, subjected to 2-D electrophoresis and stained for total proteins and phosphoproteins. The comparison of C and T gels was performed by gel image analysis for the change of normalized spot volumes. Spots with volumes equal to or exceeding C/T ratios of ±1.5 were robotically picked for in- gel digestion with trypsin and submitted for protein identification by nanoLC/MS/MS analysis. Identified proteins covered a wide range of biological activity. As kinin peptides are known to rapidly stimulate transepithelial secretion of electrolytes and water by Malpighian tubules, we focused on those proteins that might mediate the increase in transepithelial secretion. We found that AK- III reduces the cytosolic presence of subunits A and B of the V-type H+ ATPase, endoplasmin, calreticulin, annexin, type II regulatory subunit of protein kinase A (PKA) and rab GDP dissociation inhibitor and increases the cytosolic presence of adducin, actin, Ca2+-binding protein regucalcin/SMP30 and actin-depolymerizing factor.
    [Show full text]
  • Calcium-Binding Proteins Annexin A2 and S100A6 Are Sensors of Tubular Injury and Recovery in Acute Renal Failure
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Kidney International, Vol. 68 (2005), pp. 2694–2703 Calcium-binding proteins annexin A2 and S100A6 are sensors of tubular injury and recovery in acute renal failure CHAO-WEN CHENG,ABDALLA RIFAI,SHUK-MAN KA,HAO-AI SHUI,YUH-FENG LIN,WEI-HWA LEE, and ANN CHEN Graduate Institute of Life Sciences, Graduate Institute of Medical Sciences, Department of Internal Medicine, Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taiwan, Republic of China; and Department of Pathology, Rhode Island Hospital, Rhode Island Calcium-binding proteins annexin A2 and S100A6 are sensors Acute tubular necrosis is the most common pathologic of tubular injury and recovery in acute renal failure. entity responsible for the clinical state of acute renal fail- Background. Rise in cellular calcium is associated with acute ure (ARF) [1, 2]. The two main causes of acute tubu- tubular necrosis, the most common cause of acute renal failure (ARF). The mechanisms that calcium signaling induce in the lar necrosis are ischemic and toxic injuries [3]. In the quiescent tubular cells to proliferate and differentiate during latter type, a variety of renal environmental substances acute tubular necrosis have not been elucidated. that include heavy metals such as mercury, lead, and ura- Methods. Acute tubular necrosis induced in mice by sin- nium are known to cause ARF.Nephrotoxic acute tubular gle intravenous injection of uranyl nitrate and examined af- necrosis is histologically evident as epithelial cell necrosis, ter 1, 3, 7, and 14 days.
    [Show full text]
  • Annexin A2 Complexes with S100 Proteins
    British Journal of DOI:10.1111/bph.12978 www.brjpharmacol.org BJP Pharmacology Themed Section: Annexins VII Programme Correspondence Dr Lodewijk V Dekker, School of Pharmacy, Centre for REVIEW Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK. E-mail: Annexin A2 complexes [email protected] ---------------------------------------------------------------- Received with S100 proteins: 18 July 2014 Revised 16 September 2014 structure, function and Accepted 5 October 2014 pharmacological manipulation Yidong Liu, Helene K Myrvang and Lodewijk V Dekker School of Pharmacy, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK Annexin A2 (AnxA2) was originally identified as a substrate of the pp60v-src oncoprotein in transformed chicken embryonic fibroblasts. It is an abundant protein that associates with biological membranes as well as the actin cytoskeleton, and has been implicated in intracellular vesicle fusion, the organization of membrane domains, lipid rafts and membrane-cytoskeleton contacts. In addition to an intracellular role, AnxA2 has been reported to participate in processes localized to the cell surface including extracellular protease regulation and cell-cell interactions. There are many reports showing that AnxA2 is differentially expressed between normal and malignant tissue and potentially involved in tumour progression. An important aspect of AnxA2 function relates to its interaction with small Ca2+-dependent adaptor proteins called S100 proteins, which is the topic of this review. The interaction between AnxA2 and S100A10 has been very well characterized historically; more recently, other S100 proteins have been shown to interact with AnxA2 as well. The biochemical evidence for the occurrence of these protein interactions will be discussed, as well as their function.
    [Show full text]
  • Structure – Function Studies of Annexin A2 & A5
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by University of Minnesota Digital Conservancy Structure – Function Studies of Annexin A2 & A5 Stephanie Schramel Swenson College of Science and Engineering University of Minnesota Duluth [email protected] Introduction: Within the cell, proteins exist to maintain the intracellular level of calcium ions (Ca2+), allowing calcium to act as a secondary messenger. Annexins are a group of these proteins located in muscle cells which bind calcium in order to bind to phospholipids in the plasma membrane. Although there are multiple forms of this protein found in humans, all contain a similar structural basis consisting of an amino-terminal head domain, a carboxyl terminal domain, and conserved structural repeats of around 70 residues. Both the calcium binding and membrane binding sites are located at the carboxyl domain. Annexins bind to negatively charged phospholipids at the cellular membranes, which differentiates them from other calcium binding proteins. Exocytosis/endocytosis, ion transport, and transport of vesicles are also processes orchestrated by these proteins.1 Specifically, annexin A2 (AnxA2) and A5 (AnxA5) are the proteins we studied. Figure 1 shows the 3-D structure of AnxA2 with calcium bound. The question we tried to understand is the mechanism for how AnxA2 and AnxA5 function at the membrane in terms of the redistribution of lipids and the effect on membrane permeability. In order to study the annexins, we grew E. coli cells that expressed our desired proteins. We then went through multiple week- long purification processes to obtain pure protein to be used for further studies.
    [Show full text]
  • Human Induced Pluripotent Stem Cell–Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation
    BASIC RESEARCH www.jasn.org Human Induced Pluripotent Stem Cell–Derived Podocytes Mature into Vascularized Glomeruli upon Experimental Transplantation † Sazia Sharmin,* Atsuhiro Taguchi,* Yusuke Kaku,* Yasuhiro Yoshimura,* Tomoko Ohmori,* ‡ † ‡ Tetsushi Sakuma, Masashi Mukoyama, Takashi Yamamoto, Hidetake Kurihara,§ and | Ryuichi Nishinakamura* *Department of Kidney Development, Institute of Molecular Embryology and Genetics, and †Department of Nephrology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; ‡Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan; §Division of Anatomy, Juntendo University School of Medicine, Tokyo, Japan; and |Japan Science and Technology Agency, CREST, Kumamoto, Japan ABSTRACT Glomerular podocytes express proteins, such as nephrin, that constitute the slit diaphragm, thereby contributing to the filtration process in the kidney. Glomerular development has been analyzed mainly in mice, whereas analysis of human kidney development has been minimal because of limited access to embryonic kidneys. We previously reported the induction of three-dimensional primordial glomeruli from human induced pluripotent stem (iPS) cells. Here, using transcription activator–like effector nuclease-mediated homologous recombination, we generated human iPS cell lines that express green fluorescent protein (GFP) in the NPHS1 locus, which encodes nephrin, and we show that GFP expression facilitated accurate visualization of nephrin-positive podocyte formation in
    [Show full text]
  • Exophagy of Annexin A2 Via RAB11, RAB8A and RAB27A in IFN-Γ
    www.nature.com/scientificreports OPEN Exophagy of annexin A2 via RAB11, RAB8A and RAB27A in IFN-γ- stimulated lung epithelial cells Received: 23 September 2016 Ying-Da Chen1, Yi-Ting Fang2, Yi-Lin Cheng1,2, Chiou-Feng Lin3,4, Li-Jin Hsu1,3,5, Shu-Ying Accepted: 7 June 2017 Wang1,2,3, Robert Anderson3,6, Chih-Peng Chang1,2,3 & Yee-Shin Lin1,2,3 Published: xx xx xxxx Annexin A2 (ANXA2), a phospholipid-binding protein, has multiple biological functions depending on its cellular localization. We previously demonstrated that IFN-γ-triggered ANXA2 secretion is associated with exosomal release. Here, we show that IFN-γ-induced autophagy is essential for the extracellular secretion of ANXA2 in lung epithelial cells. We observed colocalization of ANXA2- containing autophagosomes with multivesicular bodies (MVBs) after IFN-γ stimulation, followed by exosomal release. IFN-γ-induced exophagic release of ANXA2 could not be observed in ATG5-silenced or mutant RAB11-expressing cells. Furthermore, knockdown of RAB8A and RAB27A, but not RAB27B, reduced IFN-γ-triggered ANXA2 secretion. Surface translocation of ANXA2 enhanced efferocytosis by epithelial cells, and inhibition of different exophagic steps, including autophagosome formation, fusion of autophagosomes with MVBs, and fusion of amphisomes with plasma membrane, reduced ANXA2- mediated efferocytosis. Our data reveal a novel route of IFN-γ-induced exophagy of ANXA2. Annexin A2 (ANXA2), a Ca2+-dependent membrane-binding protein, can distribute to the nucleus, the cytosolic membrane of organelles, as well as the inner and outer leaflets of the plasma membrane in different cell types, including macrophages, endothelium, epithelium, and tumor cells1–3.
    [Show full text]
  • Annexin A2 Expression and Partners During Epithelial Cell Differentiation Malik Zibouche, Françoise Illien, Jesus Ayala-Sanmartin
    Annexin A2 expression and partners during epithelial cell differentiation Malik Zibouche, Françoise Illien, Jesus Ayala-Sanmartin To cite this version: Malik Zibouche, Françoise Illien, Jesus Ayala-Sanmartin. Annexin A2 expression and partners during epithelial cell differentiation. Biochemistry and Cell Biology, NRC Research Press, 2019, 10.1139/bcb- 2018-0393. hal-02335473 HAL Id: hal-02335473 https://hal.archives-ouvertes.fr/hal-02335473 Submitted on 28 Oct 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Annexin A2 expression and partners during epithelial cell differentiation Malik Zibouche, Françoise Illien and Jesus Ayala-Sanmartin* CNRS, Université Sorbonne, École normale supérieure, Université PSL, Laboratoire des biomolécules, 75005 Paris, France * Corresponding author: J. Ayala-Sanmartin, CNRS, UMR 7203, Laboratoire des Biomolécules, 4 Place Jussieu, BP 182, 75252 Paris, France. Tel: 33 1 44 27 32 16. Fax: 33 1 44 27 71 50. E-mail: [email protected] Abstract The members of the annexin family of calcium- and phospholipid-binding proteins participate in different cellular processes. Annexin A2 binds to S100A10 forming a functional heterotetrameric protein that has been involved in many cellular functions such as exocytosis, endocytosis, cell junction formation and actin cytoskeleton dynamics.
    [Show full text]
  • Annexin A2 Is an Independent Prognostic Biomarker for Evaluating the Malignant Progression of Laryngeal Cancer
    EXPERIMENTAL AND THERAPEUTIC MEDICINE 14: 6113-6118, 2017 Annexin A2 is an independent prognostic biomarker for evaluating the malignant progression of laryngeal cancer SHI LUO, CHUBO XIE, PING WU, JIAN HE, YAOYUN TANG, JING XU and SUPING ZHAO Department of Otorhinolaryngology Head and Neck Surgery, Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, Hunan 410008, P.R. China Received December 16, 2016; Accepted July 7, 2017 DOI: 10.3892/etm.2017.5298 Abstract. Due to the lack of a definite diagnosis, a frequent radiotherapy; therefore, the clinical outcome for patients with recurrence rate and resistance to chemotherapy or radio- advanced laryngeal cancer remains poor (2,3). Thus, more therapy, the clinical outcome for patients with advanced effective diagnostic and therapeutic targets are required. laryngeal cancer has not improved over the last decade. Annexin family proteins promote invasion and metastasis Annexin A2 is associated with the invasion and metastasis in several types of cancer, including breast cancer, esophageal of cancer cells. In the present study, it was demonstrated carcinoma, liver cancer and nasopharyngeal carcinoma (4). using differential proteomics analysis that Annexin A2 is However, different members of the Annexin family proteins highly expressed in laryngeal carcinoma tissues and this was are either up‑ or downregulated in cancer tissue, so different confirmed using immunohistochemistry, which demonstrated Annexins may perform different roles in specific types of that the expression of Annexin A2 in laryngeal carcinoma cancer (5,6). It has been demonstrated that Annexin A1 is tissues was significantly higher than in healthy adjacent tissue. associated with esophageal cancer and is overexpressed in In addition, its potential predictive value in the prognosis of laryngeal carcinoma cells; it may therefore progressively patients with laryngeal carcinoma was evaluated.
    [Show full text]
  • Transbilayer Phospholipid Movement Facilitates the Translocation of Annexin Across Membranes Sarah E
    © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2018) 131, jcs217034. doi:10.1242/jcs.217034 RESEARCH ARTICLE Transbilayer phospholipid movement facilitates the translocation of annexin across membranes Sarah E. Stewart1,*, Avraham Ashkenazi2,*, Athena Williamson1, David C. Rubinsztein2,3,‡ and Kevin Moreau1,‡ ABSTRACT Nickel and Rabouille, 2009; Rabouille et al., 2012; Rabouille, 2017). Annexins are cytosolic phospholipid-binding proteins that can be Interestingly, all four types of secretion involve crossing a membrane. found on the outer leaflet of the plasma membrane. The extracellular For types I and II, proteins are directly translocated across the functions of annexin include modulating fibrinolysis activity and plasma membrane. In type I, protein translocation across the cell migration. Despite having well-described extracellular functions, plasma membrane is either mediated by protein complexes, pores the mechanism of annexin transport from the cytoplasmic inner leaflet or is self-mediated (unfacilitated). In type II secretion, translocation to the extracellular outer leaflet of the plasma membrane remains is mediated by ATP-binding-cassette (ABC) transporter proteins. unclear. Here, we show that the transbilayer movement of phospholipids Type III describes the secretion of cytoplasmic proteins that first enter facilitates the transport of annexins A2 and A5 across membranes in the lumen of an organelle, which then fuses with the plasma cells and in liposomes. We identified TMEM16F (also known as membrane. The type IV secretion concerns transmembrane proteins, anoctamin-6, ANO6) as a lipid scramblase required for transport of which are inserted in the ER membrane but reach the plasma these annexins to the outer leaflet of the plasma membrane.
    [Show full text]