DOCSLIB.ORG

FERM Domain Mediates Apical Targeting of PTP-BL 3301 and Incubating for 30 Minutes at Room Temperature

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
FERM Domain Mediates Apical Targeting of PTP-BL 3301 and Incubating for 30 Minutes at Room Temperature Journal of Cell Science 112, 3299-3308 (1999) 3299 Printed in Great Britain © The Company of Biologists Limited 1999 JCS0400 A FERM domain governs apical confinement of PTP-BL in epithelial cells Edwin Cuppen, Mietske Wijers, Jan Schepens, Jack Fransen, Bé Wieringa and Wiljan Hendriks* Department of Cell Biology, Institute of Cellular Signalling, University of Nijmegen, Adelbertusplein 1, 6525 EK Nijmegen, The Netherlands *Author for correspondence (e-mail: [email protected]) Accepted 2 July; published on WWW 22 September 1999 SUMMARY PTP-BL is a cytosolic multidomain protein tyrosine nor can we detect a direct interaction by phosphatase that shares homologies with several immunoprecipitation assays. Fluorescence recovery after submembranous and tumor suppressor proteins. Here we photobleaching (FRAP) experiments show that PTP-BL show, by transient expression of modular protein domains confinement is based on a dynamic steady state and that of PTP-BL in epithelial MDCK cells, that the presence of complete redistribution of the protein may occur within 20 a FERM domain in the protein is both necessary and minutes. Our observations suggest that relocation is sufficient for its targeting to the apical side of epithelial mediated via a cytosolic pool, rather than by lateral cells. Furthermore, immuno-electron microscopy on stable movement. Finally, we show that PTP-BL phosphatase expressing MDCK pools, that were obtained using an domains are involved in homotypic interactions, as EGFP-based cell sorting protocol, revealed that FERM demonstrated by yeast two-hybrid assays. Both the highly domain containing fusion proteins are enriched in restricted subcellular compartmentalization and its specific microvilli and have a typical submembranous location associative properties may provide the appropriate at about 10-15 nm from the plasma membrane. conditions for regulating substrate specificity and catalytic Immunofluorescence microscopy suggested colocalization activity of this member of the PTP family. of the FERM domain moiety with the membrane- cytoskeleton linker ezrin. However, at the electron Key words: Polarized, Band 4.1, PDZ, Immuno-electron microscopy, microscopy level this colocalization cannot be confirmed Green fluorescent protein, EGFP, Protein tyrosine phosphatase INTRODUCTION receptor-type RPTPα gives structural support for a model in which enzyme inhibition is the consequence of dimerization, Reversible phosphorylation of proteins on tyrosine residues is by showing that the active site cleft in the subunits is blocked a key event in the regulation of various cellular processes, such by protruding loops of the opposing domain (Bilwes et al., as proliferation, differentiation, cell motility, cell-cell 1996). Structural data on the RPTPµ membrane proximal PTP interactions, metabolism, gene transcription, and the immune domain, however, show that this model might not be response (Hunter, 1996). Proper initiation, propagation and universally applicable (Hoffmann et al., 1997). Also, for the termination of the signaling cascades involved are thus cytosolic PTP SHP-2, for example, it was shown that N- critically dependent on the mode of action of individual terminal SH2 domains are involved in blocking its catalytic site members of the large families of protein tyrosine kinases in an intramolecular interaction (Hof et al., 1998). (PTKs) and protein tyrosine phosphatases (PTPs) (Hunter, Importantly, also targeting of the enzyme to highly restricted 1998; Ninfa and Dixon, 1994). It is now well established that subcellular micro-compartments may control PTP activity and receptor PTKs are activated by an intermolecular mechanism substrate specificity. In this light, the existence of a variety of upon ligand-induced dimerization (Lemmon and Schlessinger, modular protein domains in cytosolic PTPs, that may function 1994). Cytoplasmic PTKs, on the other hand, are activated by as ‘zip codes’ by directing the protein to the correct cellular both intra- and intermolecular mechanisms (Thomas and ‘address’ (Mauro and Dixon, 1994), is intriguing. Candidate Brugge, 1997). Much less is known regarding the processes domains that may regulate activity and specificity are that antagonize the effects of PTKs or, for that matter, the particularly abundant in the large cytosolic protein tyrosine functional activation of PTPs. Recent data suggest that for phosphatase PTP-BL (Hendriks et al., 1995) or RIP (Chida et PTPs similar mechanisms as for PTKs do exist (reviewed by al., 1995), which is the mouse homologue of human PTP- Weiss and Schlessinger, 1998). For example, ligand-induced BAS/PTPL1/PTP1E/FAP-1 (Banville et al., 1994; Maekawa et dimerization of an EGFR-CD45 chimera was found to inhibit al., 1994; Saras et al., 1994; Sato et al., 1995). The PTP activity and TCR signaling (Majeti et al., 1998). The approximately 250 kDa protein, that is expressed mainly in crystal structure of the membrane-proximal PTP domain of the epithelia (Hendriks et al., 1995; Cuppen et al., 1998; Thomas 3300 E. Cuppen and others et al., 1998), harbors a large N-terminal domain with no interaction of the catalytic PTP-BL phosphatase domain. We obvious homology to other proteins except for a potential suggest that subcellular targeting of PTP-BL to a highly leucine zipper (Saras et al., 1994), followed by a FERM restricted microcompartment within the cell, possibly together domain, five PDZ motifs and a C-terminal PTP domain (see with a dimerization-induced shift in activity, may be an also Fig. 2). Outside its natural context, the PTP-BL important determinant in confining its specificity and activity. phosphatase domain has a high rate of activity with a rather promiscuous substrate specificity (Cuppen et al., 1998; Hendriks et al., 1995). For example, a PTP-BL phosphatase MATERIALS AND METHODS domain-GST fusion protein is able to completely eliminate the extensive phosphorylation of endogenous proteins by the Expression plasmid constructions activated tyrosine kinase present in Escherichia coli TKX-1 The cDNA restriction fragments encoding segments of PTP-BL cells (E. Cuppen, unpublished observations). Obviously, a (GenBank accession number Z32740) as indicated in Fig. 2 were regulatory mechanism must be involved in imposing the proper subcloned in pEGFP vectors (Clontech) or in modified versions of the restrictions on PTP-BL’s catalytic activity at endogenous sites. eukaryotic expression vector pSG5 (Green et al., 1988). In the pSG5 The modular domains, like the PDZ and FERM motifs that are vector we have introduced an in-frame VSV-G epitope tag sequence present in PTP-BL, may provide this function in vivo. to allow the production of fusion-proteins that can be detected with the α-VSV monoclonal antibody P5D4. Furthermore, we introduced PDZ motifs are small protein modules that mediate protein- either an initiator AUG codon or a stop codon for proper start or protein interactions (Fanning and Anderson, 1996; termination of translation (see also Cuppen et al., 1998). BL-PDZ-I- Ranganathan and Ross, 1997; Saras and Heldin, 1996) and can V and BL-P were tagged at their N-terminal end, whereas BL-FERM associate with transmembrane proteins (Sheng, 1996), is tagged at its C terminus. A single point mutation at amino acid cytoskeletal components (Xia et al., 1997) and signal 2344, aspartate to alanine, was introduced in BL-P by PCR using transduction enzymes (Ranganathan and Ross, 1997). Indeed, synthetic oligonucleotides, resulting in the catalytically inactive several proteins that interact with PTP-BL PDZ motifs have mutant BL-P-DA. The BL-Nter and the BL-Nter-FERM constructs been identified (Cuppen et al., 1998; Saras et al., 1997; E. were tagged C-terminally with EGFP by subcloning into the pEGFP- Cuppen, unpublished observations), but thus far no clues as to N3 vector (Clontech), whereas BL-FERM+PDZ-I-V and EGFP-BL- explain PTP-BL’s subcellular localization in vivo have been FERM were tagged N-terminally by subcloning into the pEGFP-C1 vector (Clontech). BL-FERM+PDZ-I-V+P was expressed as an obtained. untagged protein from the pSG5 vector. Full length PTP-BL cDNA, The FERM domain in PTP-BL is a segment commonly including its 3′-UTR, was inserted into pEGFP-N3, allowing the found in a family of peripheral membrane proteins that production of untagged PTP-BL (BL-FL). Subsequently, this function as membrane-cytoskeleton linkers (Chishti et al., construct was modified by PCR to generate a PTP-BL-EGFP fusion 1998). This family includes erythrocyte protein 4.1, ezrin, construct (BL-FL-EGFP). To this end, we introduced a XhoI site at radixin and moesin, but also the tumor suppressors Expanded the position of the stop codon, thereby removing the 3′-UTR including and merlin and the protein tyrosine phosphatases PTP-MEG, the stop codon, before introducing the EGFP moiety. Detailed cloning PTPH1, PTPD1 and CDEP. The FERM domain in erythrocyte information can be obtained from the authors upon request. protein 4.1 binds a wide variety of molecules including ATP, Antibodies PIP2, phosphatidylserine, calmodulin, p55, glycophorin A, The affinity-purified polyclonal antisera α-BL-PDZ-I and α-BL-P, glycophorin C, Band 3 and CD44 (see Chishti et al., 1998 for directed against the first PDZ motif and the phosphatase domain of references). Ezrin, radixin and moesin (ERMs) also function PTP-BL, respectively, and the monoclonal α-VSV antibody P5D4 as molecular linkers that connect cell-surface transmembrane have been described elsewhere (Cuppen et al., 1998; Kreis, 1986). The proteins to the
Load more

Recommended publications
  • Myosin Myth4-FERM Structures Highlight Important Principles of Convergent Evolution
    Myosin MyTH4-FERM structures highlight important principles of convergent evolution Vicente José Planelles-Herreroa,b, Florian Blanca,c, Serena Sirigua, Helena Sirkiaa, Jeffrey Clausea, Yannick Souriguesa, Daniel O. Johnsrudd, Beatrice Amiguesa, Marco Cecchinic, Susan P. Gilberte, Anne Houdussea,1,2, and Margaret A. Titusd,1,2 aStructural Motility, Institut Curie, CNRS, UMR 144, PSL Research University, F-75005 Paris, France; bUPMC Université de Paris 6, Institut de Formation Doctorale, Sorbonne Universités, 75252 Paris Cedex 05, France; cLaboratoire d’Ingénierie des Fonctions Moléculaires, Institut de Science et d’Ingénierie Supramoléculaires, UMR 7006 CNRS, Université de Strasbourg, F-67083 Strasbourg Cedex, France; dDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455; and eDepartment of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY 12180 Edited by James A. Spudich, Stanford University School of Medicine, Stanford, CA, and approved March 31, 2016 (received for review January 15, 2016) Myosins containing MyTH4-FERM (myosin tail homology 4-band (Fig. 1). These MF myosins are widespread and likely quite an- 4.1, ezrin, radixin, moesin, or MF) domains in their tails are found cient because they are found in many different branches of the in a wide range of phylogenetically divergent organisms, such as phylogenetic tree (5, 6), including Opisthokonts (which includes humans and the social amoeba Dictyostelium (Dd). Interestingly, Metazoa, unicellular Holozoa, and Fungi), Amoebozoa, and the evolutionarily distant MF myosins have similar roles in the exten- SAR (Stramenopiles, Alveolates, and Rhizaria) (Fig. 1 A and B). sion of actin-filled membrane protrusions such as filopodia and Over the course of hundreds of millions years of parallel evolution bind to microtubules (MT), suggesting that the core functions of the MF myosins have acquired or maintained roles in the formation these MF myosins have been highly conserved over evolution.
    [Show full text]
  • Myth4 and FERM Have Overlapping and Distinct Roles in the Function of Myo1, a Class XIV Myosin in Tetrahymena Thermophila
    City University of New York (CUNY) CUNY Academic Works All Dissertations, Theses, and Capstone Projects Dissertations, Theses, and Capstone Projects 2011 MyTH4 and FERM Have Overlapping and Distinct Roles in the Function of Myo1, a Class XIV Myosin in Tetrahymena thermophila Michael Gotesman Graduate Center, City University of New York How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/gc_etds/1643 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] MyTH4 and FERM Have Overlapping and Distinct Roles in the Function of Myo1, a Class XIV Myosin in Tetrahymena thermophila By Michael Gotesman A dissertation submitted to the Graduate Faculty in Biology in partial fulfillment of the requirements for the degree of Doctor of Philosophy, The City University of New York 2011 This manuscript has been read and accepted for the Graduate Faculty in Biology in satisfaction of the dissertation requirements for the degree of Doctor of Philosophy. _____________ ________________________________________________ Date Chair of Examining Committee Dr. Ray H. Gavin, Brooklyn College _____________ ________________________________________________ Date Executive Officer Dr. Laurel A. Eckhardt ________________________________________________ Dr. Shaneen M. Singh, Brooklyn College ________________________________________________ Dr. Theodore R. Muth, Brooklyn College ________________________________________________ Dr. Chang-Hui Shen, College of Staten Island ________________________________________________ Dr. Selwyn A. Williams, New York City of Technology ________________________________________________ Dr. Christina King-Smith, Saint Joseph’s University Supervising Committee The City University of New York ii Abstract MyTH4 and FERM Have Overlapping and Distinct Roles in the Function of Myo1, a Class XIV Myosin in Tetrahymena thermophila By Michael Gotesman Adviser: Dr.
    [Show full text]
  • FRNK Regulatory Complex Formation with FAK Is Regulated by ERK Mediated Serine 217 Phosphorylation
    Loyola University Chicago Loyola eCommons Dissertations Theses and Dissertations 2017 FRNK Regulatory Complex Formation with FAK Is Regulated by ERK Mediated Serine 217 Phosphorylation Taylor J. Zak Loyola University Chicago Follow this and additional works at: https://ecommons.luc.edu/luc_diss Part of the Biochemistry, Biophysics, and Structural Biology Commons Recommended Citation Zak, Taylor J., "FRNK Regulatory Complex Formation with FAK Is Regulated by ERK Mediated Serine 217 Phosphorylation" (2017). Dissertations. 2604. https://ecommons.luc.edu/luc_diss/2604 This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion in Dissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. Copyright © 2017 Taylor J. Zak LOYOLA UNIVERSITY CHICAGO FRNK REGULATORY COMPLEX FORMATION WITH FAK IS REGUALTED BY ERK MEDIATED SERINE 217 PHOSPHORYLATION A DISSERTATION SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY PROGRAM IN CELL AND MOLECULAR PHYSIOLOGY BY TAYLOR J. ZAK CHICAGO, ILLINOIS MAY 2017 Copyright by Taylor J. Zak, 2017 All rights reserved. Dedicated to my wife Stacey ACKNOWLEDGEMENTS This dissertation would not be possible without the day to day guidance of doctors Seth Robia and Allen Samarel. Dr. Samarel’s guidance was missed during the final year of my dissertation as he transitioned to an emeritus professor and I am forever grateful to Dr. Robia for taking on some of Dr. Samarel’s role.
    [Show full text]
  • ERM Protein Family
    Cell Biology 2018; 6(2): 20-32 http://www.sciencepublishinggroup.com/j/cb doi: 10.11648/j.cb.20180602.11 ISSN: 2330-0175 (Print); ISSN: 2330-0183 (Online) Structure and Functions: ERM Protein Family Divine Mensah Sedzro 1, †, Sm Faysal Bellah 1, 2, †, *, Hameed Akbar 1, Sardar Mohammad Saker Billah 3 1Laboratory of Cellular Dynamics, School of Life Science, University of Science and Technology of China, Hefei, China 2Department of Pharmacy, Manarat International University, Dhaka, Bangladesh 3Department of Chemistry, Govt. M. M. University College, Jessore, Bangladesh Email address: *Corresponding author † These authors contributed equally to this work To cite this article: Divine Mensah Sedzro, Sm Faysal Bellah, Hameed Akbar, Sardar Mohammad Saker Billah. Structure and Functions: ERM Protein Family. Cell Biology . Vol. 6, No. 2, 2018, pp. 20-32. doi: 10.11648/j.cb.20180602.11 Received : September 15, 2018; Accepted : October 6, 2018; Published : October 29, 2018 Abstract: Preservation of the structural integrity of the cell depends on the plasma membrane in eukaryotic cells. Interaction between plasma membrane, cytoskeleton and proper anchorage influence regular cellular processes. The needed regulated connection between the membrane and the underlying actin cytoskeleton is therefore made available by the ERM (Ezrin, Radixin, and Moesin) family of proteins. ERM proteins also afford the required environment for the diffusion of signals in reactions to extracellular signals. Other studies have confirmed the importance of ERM proteins in different mode organisms and in cultured cells to emphasize the generation and maintenance of specific domains of the plasma membrane. An essential attribute of almost all cells are the specialized membrane domains.
    [Show full text]
  • A Novel FERM Domain Including Guanine Nucleotide Exchange Factor Is Involved in Rac Signaling and Regulates Neurite Remodeling
    The Journal of Neuroscience, October 1, 2002, 22(19):8504–8513 A Novel FERM Domain Including Guanine Nucleotide Exchange Factor Is Involved in Rac Signaling and Regulates Neurite Remodeling Tateki Kubo,1,2,3 Toshihide Yamashita,1,3 Atsushi Yamaguchi,1,3 Hideki Sumimoto,4 Ko Hosokawa,2 and Masaya Tohyama1,3 Departments of 1Anatomy and Neuroscience and 2Plastic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan, 3Core Research for Evolutional Science and Technology of Japan Science and Technology Corporation, Kawaguchi, Saitama, 332-0012, Japan, and 4Medical Institute of Bioregulation, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan The Rho family of small GTPases, key regulators of the actin testis, as well as embryonic hippocampal and cortical neurons. cytoskeleton in eukaryotic cells from yeast to human, is impli- FIR was found to activate the biochemical pathway specific for cated in the control of neuronal morphology. Guanine nucleo- Rac1 but not for RhoA or Cdc42. Ectopic expression of FIR in tide exchange factors (GEFs) are upstream positive regulators the cortical neurons resulted in significantly shortened neurites of Rho GTPases and integrate extracellular signaling for appro- and excessive growth cones, presumably mediated by Rac1. priate activation of Rho GTPases at specific subcellular re- These results suggest that FIR may regulate neurite remodeling gions. Here we describe the identification of a novel Dbl family by mediating the signaling pathways from membrane proteins GEF for Rho GTPases in Homo sapiens and Mus musculus.It to Rac. contains a tandem Dbl homology–pleckstrin homology domain and FERM domain, characteristic of the plasma membrane Key words: Rac1; Rho guanine nucleotide exchange factor; proteins linker.
    [Show full text]
  • FAK in Cancer: Mechanistic Findings and Clinical Applications
    REVIEWS FAK in cancer: mechanistic findings and clinical applications Florian J. Sulzmaier, Christine Jean and David D. Schlaepfer Abstract | Focal adhesion kinase (FAK) is a cytoplasmic protein tyrosine kinase that is overexpressed and activated in several advanced-stage solid cancers. FAK promotes tumour progression and metastasis through effects on cancer cells, as well as stromal cells of the tumour microenvironment. The kinase-dependent and kinase-independent functions of FAK control cell movement, invasion, survival, gene expression and cancer stem cell self-renewal. Small molecule FAK inhibitors decrease tumour growth and metastasis in several preclinical models and have initial clinical activity in patients with limited adverse events. In this Review, we discuss FAK signalling effects on both tumour and stromal cell biology that provide rationale and support for future therapeutic opportunities. Focal Adhesion Kinase (FAK) is a multifunctional regu- models that have been used to elucidate new roles for lator of cell signalling within the tumour microenviron- FAK in endothelial cells (ECs) and discuss how stromal ment1–3. During development and in various tumours, FAK signalling contributes to tumour progression. FAK promotes cell motility, survival and proliferation Finally, we summarize new translational developments through kinase-dependent and kinase-independent using small molecule FAK inhibitors. mechanisms. In the past few years, Phase I and II clinical trials have been initiated with FAK inhibitors; however, FAK regulation some of the functions of FAK in tumorigenesis remain Control of FAK expression. Nuclear factor-κB (NF‑κB) under investigation. and p53 are well-characterized transcription factors that Chromosomal region 8q24.3, which encompasses activate and repress the PTK2 promoter, respectively10,11.
    [Show full text]
  • Molecular Interactions of Talin and Actin: a Study on the Specific Domains of Talin Involved in the Interactions Ho-Sup Lee Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2002 Molecular interactions of talin and actin: a study on the specific domains of talin involved in the interactions Ho-Sup Lee Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biochemistry Commons, Cell Biology Commons, and the Molecular Biology Commons Recommended Citation Lee, Ho-Sup, "Molecular interactions of talin and actin: a study on the specific domains of talin involved in the interactions " (2002). Retrospective Theses and Dissertations. 528. https://lib.dr.iastate.edu/rtd/528 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion.
    [Show full text]
  • Emergence and Evolution of ERM Proteins and Merlin in Metazoans
    GBE Emergence and Evolution of ERM Proteins and Merlin in Metazoans Victoria Shabardina1,*, Yukie Kashima2, Yutaka Suzuki3, and Wojciech Makalowski1 1Institue of Bioinformatics, University of Muenster, Germany 2Department of Computational Biology and Medical Sciences, The University of Tokyo, Kashiwa, Japan 3Laboratory of Systems Genomics, Department of Computational Biology and Medical Sciences, The University of Tokyo, Kashiwa, Japan *Corresponding author: E-mail: [email protected]. Accepted: December 2, 2019 Abstract Ezrin, radixin, moesin, and merlin are cytoskeletal proteins, whose functions are specific to metazoans. They participate in cell cortex rearrangement, including cell–cell contact formation, and play an important role in cancer progression. Here, we have performed a comprehensive phylogenetic analysis of the proteins spanning 87 species. The results describe a possible mechanism for the protein family origin in the root of Metazoa, paralogs diversification in vertebrates, and acquisition of novel functions, including tumor suppression. In addition, a merlin paralog, present in most vertebrates but lost in mammals, has been described here for the first time. We have also highlighted a set of amino acid variations within the conserved motifs as the candidates for determining physiological differences between ERM paralogs. Key words: protein evolution, paralogs fate, ERM phylogeny. Introduction phosphorylation of a conserved threonine in the CERMAD Ezrin, radixin, and moesin of the ERM protein family, further (Yonemura et al. 2002; Niggli and Rossy 2008). An important ERMs, are cytoskeleton proteins that mediate physical con- role during this transition belongs to the middle a-helical do- nection between intermembrane proteins and actin filaments main. In the dormant ERMs, it forms a coiled-coil structure, (Bretscher et al.
    [Show full text]
  • Membrane Targeting of Protein Tyrosine Phosphatase PTPL1 Through Its FERM Domain Via Binding to Phosphatidylinositol 4,5-Biphosphate
    Membrane targeting of protein tyrosine phosphatase PTPL1 through its FERM domain via binding to phosphatidylinositol 4,5-biphosphate. Guillaume Bompard, Marianne Martin, Christian Roy, Françoise Vignon, Gilles Freiss To cite this version: Guillaume Bompard, Marianne Martin, Christian Roy, Françoise Vignon, Gilles Freiss. Membrane targeting of protein tyrosine phosphatase PTPL1 through its FERM domain via binding to phos- phatidylinositol 4,5-biphosphate.. Journal of Cell Science, Company of Biologists, 2003, 116 (Pt 12), pp.2519-30. 10.1242/jcs.00448. inserm-00150999 HAL Id: inserm-00150999 https://www.hal.inserm.fr/inserm-00150999 Submitted on 19 Nov 2009 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. Membrane targeting of protein tyrosine phosphatase PTPL1 through its FERM domain via binding to phosphatidylinositol 4,5-biphosphate Key words: Protein tyrosine phosphatase, FERM domain, PI(4,5)P2-binding sites, neomycin, apical localization. Guillaume Bompard1,*, Marianne Martin2, Christian Roy2, Françoise Vignon1 and Gilles Freiss1. 1Inserm U540, Endocrinologie Moléculaire et Cellulaire des Cancers, Montpellier, France and 2Dynamique Moléculaire des Interactions Membranaires, Université Montpellier II, Unité Mixte de Recherche (UMR) CNRS 5539, Montpellier Cedex 5, France. *Author for correspondence (present address: School of Biosciences, University of Birmingham, UK.
    [Show full text]
  • Evolutionary Stories Told by One Protein Family: ERM Phylogeny in Metazoans
    bioRxiv preprint doi: https://doi.org/10.1101/631770; this version posted May 9, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. Evolutionary stories told by one protein family: ERM phylogeny in metazoans Shabardina V.1, Kashima Y.2, Suzuki Y.2, Makalowski W.1 1Institue of Bioinformatics, University of Muenster, Niels-Stensen-Strasse 14, Muenster, 48149, Germany. 2Laboratory of Systems Genomics, Department of Computational Biology and Medical Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba, 277-8562, Japan. Abstract Ezrin, radixin, moesin, and merlin are the cytoskeletal proteins that participate in cell cortex rearrangements and also play role in cancer progression. Here we perform a comprehensive phylogenetic analysis of the protein family in metazoans spanning 87 species. The results describe a possible mechanism of the proteins origin in the root of Metazoa, paralogs diversification in vertebrates and acquirement of novel functions, including tumor suppression. In addition, a merlin paralog, present in most of vertebrates, but lost in mammals, has been described. We also highlight the set of amino acid variations within the conserved motifs as the candidates for determining physiological differences between the ERM protein paralogs. Introduction Ezrin, radixin and moesin of the ERM protein family, further ERMs, are cytoskeleton proteins that mediate physical connection between intermembrane proteins and actin filaments (Bretscher, Edwards and Fehon, 2002). They also act as signaling molecules, for example, as intermediaries in Rho signaling (Ivetic and 1 bioRxiv preprint doi: https://doi.org/10.1101/631770; this version posted May 9, 2019.
    [Show full text]
  • Crystal Structure of the FERM-Folded Talin Head Reveals the Determinants for Integrin Binding
    Crystal structure of the FERM-folded talin head reveals the determinants for integrin binding Pingfeng Zhanga,1, Latifeh Azizib,c, Sampo Kukkurainenb,c,2, Tong Gaoa,2, Mo Baikoghlid,2, Marie-Claude Jacquiere,2, Yijuan Suna, Juha A. E. Määttäb,c, R. Holland Chengd, Bernhard Wehrle-Hallere,3, Vesa P. Hytönenb,c,3, and Jinhua Wua,3 aMolecular Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA 19111; bFaculty of Medicine and Health Technology, Tampere University, FI-33520 Tampere, Finland; cDepartment of Clinical Chemistry, Fimlab Laboratories, FI-33520 Tampere, Finland; dDepartment of Molecular and Cellular Biology, University of California, Davis, CA 95616; and eDepartment of Cell Physiology and Metabolism, Centre Médical Universitaire, University of Geneva, 1211 Geneva 4, Switzerland Edited by Janet L. Smith, University of Michigan–Ann Arbor, Ann Arbor, MI, and approved October 28, 2020 (received for review July 10, 2020) Binding of the intracellular adapter proteins talin and its cofactor, and F3) that have been shown by X-ray crystallography for sev- kindlin, to the integrin receptors induces integrin activation and eral members of the family, including kindlin-2, a coactivator of clustering. These processes are essential for cell adhesion, migra- integrin and most homologous to talin (14). Interestingly, unlike tion, and organ development. Although the talin head, the a typical FERM domain, a functionally impaired 1 to 400 talin integrin-binding segment in talin, possesses a typical FERM-do- fragment, missing the loop in the F1 subdomain (del139–168), as main sequence, a truncated form has been crystallized in an un- well as the C-terminal poly-lysine motif in the F3 domain, has expected, elongated form.
    [Show full text]
  • PDF Printing 600
    BIO 5440 Burr, 2012 I. Non-muscle Actin: Bundles and Networks 1. Actin bundles: (“Plus” ends are attached to membranes) a) “Tight bundles”: bundled by fascin [fibroblasts], or villin, fimbrin [microvilli of intestinal epithelial cells: fig 17-4, p716]. (Protein structures listed in figure 17-18, p 729) Examples: fibroblasts: lamellipodia, microspikes (10 µm), filopodia (= long microspikes: 50µm) epithelial cells: Microvilli [ fig 17-4, p716] b) “Loose bundles” (contractile): bundled by α-actinin [leaves room in between parallel actin filaments for the insertion of myosin I or II] Examples: (i) fibroblast “stress fibers” (fig 19-32, p832) ( ⇒ focal adhesions/“3D” adhesions) (fig 19-33, p834; fibronectin (Fn) Fn receptor (an Integrin) Focal adhesion kinase (fak) pp. 10-12 of handout Src kinase vinculin, talin, (ii) contractile ring in mitotic cells (fig 17-34, p742) (iii) adherens belt in epithelial cells (fig 17-4, p716) 2. Actin networks: filamin [crosslinks f-actin, leading to gel formation] (fig 17-18, p 729), versus gelsolin [Ca++-activated severing of f-actin, leading to sol formation] Examples: cell cortex; gel-sol conversions in amoeba pseudopod (Tom Stossel, American Scientist, 78, 408-423 [1990]: pp 17a,17b of handout) 3. Actin filaments are linked to membrane proteins Examples: dystrophin; the actin-spectrin network underlying the red blood cell membrane II. Polymerization Kinetics of Actin, in vitro. (p24 of handout) 1. Critical concentration for polymerization (Cc) 2. Plus and minus ends 3. Significance of ATP hydrolysis: Cc for plus end can be different from Cc for minus end 4. Treadmilling 5. Drugs affecting polymerization/depolymerization of actin: cytochalasin (destabilizes) & phalloidin (stabilizes) BIO 5440 J.G.
    [Show full text]