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PALLD Regulates Phagocytosis by Enabling Timely Actin Polymerization and Depolymerization

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PALLD Regulates Phagocytosis by Enabling Timely Actin Polymerization and Depolymerization PALLD Regulates Phagocytosis by Enabling Timely Actin Polymerization and Depolymerization This information is current as Hai-Min Sun, Xin-Lei Chen, Xin-Jie Chen, Jin Liu, Lie Ma, of September 29, 2021. Hai-Yan Wu, Qiu-Hua Huang, Xiao-Dong Xi, Tong Yin, Jiang Zhu, Zhu Chen and Sai-Juan Chen J Immunol 2017; 199:1817-1826; Prepublished online 24 July 2017; doi: 10.4049/jimmunol.1602018 Downloaded from http://www.jimmunol.org/content/199/5/1817 Supplementary http://www.jimmunol.org/content/suppl/2017/07/23/jimmunol.160201 Material 8.DCSupplemental http://www.jimmunol.org/ References This article cites 42 articles, 16 of which you can access for free at: http://www.jimmunol.org/content/199/5/1817.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 29, 2021 • No Triage! Every submission reviewed by practicing scientists • 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 © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology PALLD Regulates Phagocytosis by Enabling Timely Actin Polymerization and Depolymerization Hai-Min Sun,1 Xin-Lei Chen,1 Xin-Jie Chen,1 Jin Liu, Lie Ma, Hai-Yan Wu, Qiu-Hua Huang, Xiao-Dong Xi, Tong Yin, Jiang Zhu, Zhu Chen, and Sai-Juan Chen PALLD is an actin cross-linker supporting cellular mechanical tension. However, its involvement in the regulation of phagocytosis, a cellular activity essential for innate immunity and physiological tissue turnover, is unclear. We report that PALLD is highly induced along with all-trans-retinoic acid–induced maturation of myeloid leukemia cells, to promote Ig- or complement-opsonized phago- cytosis. PALLD mechanistically facilitates phagocytic receptor clustering by regulating actin polymerization and c-Src dynamic activation during particle binding and early phagosome formation. PALLD is also required at the nascent phagosome to recruit phosphatase oculocerebrorenal syndrome of Lowe, which regulates phosphatidylinositol-4,5-bisphosphate hydrolysis and actin depolymerization to complete phagosome closure. Collectively, our results show a new function for PALLD as a crucial regulator Downloaded from of the early phase of phagocytosis by elaborating dynamic actin polymerization and depolymerization. The Journal of Immu- nology, 2017, 199: 1817–1826. ALLD is an actin-associated protein and crucial to Mature myeloid cells maintain tissue homeostasis, repair, and establishing cellular morphology and maintaining cyto- remodeling, as well as eliminating foreign materials or pathogens P skeletal organization in various cell types (1, 2). A single through the specialized function of phagocytosis (18, 19). Phago- http://www.jimmunol.org/ PALLD gene gives rise to several isoforms, some of which are cytosis is the receptor-mediated engulfment of large particles expressed in tissue-specific patterns. The most common PALLD (diameter $0.5 mm) by plasma membrane–derived vacuoles isoform is 90–92 kDa and contains a proline-rich domain in the called phagosomes (20, 21). Phagocytosis begins as phagocytic N-terminal and three C-terminal IgC2 domains (third, fourth, and receptors are bound with certain signal moieties, such as certain fifth IgC2 domain) (3). PALLD interacts with proteins involved in intrinsic components, IgG, or complement opsonins, on the sur- actin polymerization and cross-linking, acting as a molecular scaf- face of particles. The occupied phagocytic receptors then trigger a folding protein that links proteins with different functional modali- series of complex intracellular signaling events to orchestrate ef- ties into large complexes (3–15). ficient particle internalization. The details of this process vary by guest on September 29, 2021 PALLD is absent or expressed at very low levels in peripheral depending on the substrate being internalized and receptor types blood monocytes, but is a constitutively expressed factor in nu- involved, but the clustering of occupied receptors to enhance merous types of vertebrate tissue cells. PALLD is significantly substrate-binding affinity in the phagosome is a common feature upregulated during the differentiation of monocytes into dendritic (20, 22). cells (16). PALLD upregulation during all-trans-retinoic acid Phosphatidylinositol-4,5-bisphosphate [PI(4,5)P ] is essential for (ATRA)–induced myeloid differentiation of acute promyelocytic 2 the initial actin polymerization that drives pseudopod formation, leukemia cells implies that PALLD has a role in the differentiation whereas its hydrolysis is a prerequisite for actin depolymerization that and/or function of mature myeloid cells (17). allows complete phagosome closure. In mammalian cells, PI(4,5)P2 is metabolized by conversion to PI(3,4,5)P3 via class I PI3Ks; cleavaged into inositol 3,4,5-trisphosphate and diacylglycerol by State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Rui- phospholipase C; and dephosphorylated by inositol phosphatases Jin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shang- hai 200025, China such as inositol polyphosphate-5-phosphatase and/or oculocere- 1H.-M.S., X.-L.C., and X.-J.C. contributed equally to this work. brorenal syndrome of Lowe (OCRL) (23, 24). Efficient phagocytosis relies on the focal exocytosis of intra- Received for publication December 2, 2016. Accepted for publication June 12, 2017. cellular compartments that contributes to the release of membrane This work was supported by National Key Basic Research Program of China Grant 2013CB966800, Ministry of Health Grant 201202003, Mega-Projects of Scientific tension, allowing efficient phagosome formation around large par- Research for the 12th Five-Year Plan (2013ZX09303302), National Natural Science ticles (25, 26). Phagosomes undergo a maturation process following Foundation of China Grant 81123005, and the Samuel Waxman Cancer Research Foundation Co-Principal Investigator Program. scission from the plasma membrane, sequentially fusing with endo- somes and lysosomes, and ultimately becoming phagolysosomes, Address correspondence and reprint requests to Prof. Sai-Juan Chen, Prof. Zhu Chen, or Prof. Jiang Zhu, State Key Laboratory of Medical Genomics, Shanghai Institute of which are highly acidic and hydrolase-rich organelles that degrade Hematology, RuiJin Hospital Affiliated to Shanghai Jiao Tong University School of internalized particles (20). Medicine, 197 RuiJin Road II, Shanghai 200025, China. E-mail address: sjchen@stn. sh.cn (S.-J.C.), [email protected] (Z.C.), or [email protected] (J.Z.) In this study, we demonstrate a new function for PALLD in the The online version of this article contains supplemental material. promotion of phagocytic cup extension and closure. Our data identified PALLD as a crucial factor involved in the regulation of Abbreviations used in this article: ATRA, all-trans-retinoic acid; BM, bone marrow; BMDC, BM-derived dendritic cell; KD, knockdown; NC, negative control; OCRL, F-actin dynamics throughout multiple steps of phagocytosis, in- oculocerebrorenal syndrome of Lowe; PI(4,5)P2, phosphatidylinositol-4,5-bisphosphate; cluding serum-mediated particle binding via modulation of FcgRIIA sh, short hairpin; siRNA, small interfering RNA. clustering, and delivery of PI(4,5)P2 phosphatase OCRL in nascent Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 phagosomes. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1602018 1818 PALLD REGULATES ACTIN DYNAMICS IN PHAGOCYTOSIS Materials and Methods Fluor 546 phalloidin and Alexa Fluor 635 phalloidin (Invitrogen). Src Reagents and cell culture family kinase inhibitor PP2 was purchased from Selleckchem; cytochalasin D and ATRA were purchased from Sigma-Aldrich. In addition, 1.75 mm The following Abs were used in the experiments: anti-PALLD (Proteintech Fluoresbrite Bright Blue Microspheres were purchased from Poly- and Novus Biologicals); anti-FLAG M2 and anti–b-actin (Sigma-Aldrich); sciences; unlabeled zymosan and fluorescein-conjugated zymosan were anti-FcgIIA IV.3 (Stem Cell); anti-Src (Cell Signaling and Proteintech); purchased from Invitrogen. Murine GM-CSF and IL-4 were purchased anti-OCRL (Cell Signaling and Abcam); anti-PI(4,5)P2, anti-ARP3, and from PeproTech. anti-GFP (Abcam); anti-RAC1 and anti-CDC42 (Thermo Fisher); anti– The leukemia cell lines used in this study included human acute mye- phospho-Src Tyr416, anti-EEA1, anti-RAB7, and anti-LAMP1 (Cell Sig- loblastic leukemia HL60, SKNO-1, and Kasumi-1; human acute promyelocytic naling); anti–Gr-1-PE, CD11c-FITC, and anti–CD16-PE (eBioscience); leukemia NB4; human acute myelomonocytic leukemia OCI-AML3; human CD11b-PE (BioLegend); anti-CD18 (Proteintech); Alexa Fluor 488 anti- acute monocytic leukemia U937 and THP1; and human acute erythroid leu- rabbit IgG and Alexa Fluor 594 anti-mouse IgG (Jackson); and Alexa kemia HEL. The cell lines were cultured in RPMI 1640 medium containing Downloaded from http://www.jimmunol.org/
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