DOCSLIB.ORG

Hras Intracellular Trafficking and Signal Transduction Jodi Ho-Jung Mckay Iowa State University

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Hras Intracellular Trafficking and Signal Transduction Jodi Ho-Jung Mckay Iowa State University Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2007 HRas intracellular trafficking and signal transduction Jodi Ho-Jung McKay Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biological Phenomena, Cell Phenomena, and Immunity Commons, Cancer Biology Commons, Cell Biology Commons, Genetics and Genomics Commons, and the Medical Cell Biology Commons Recommended Citation McKay, Jodi Ho-Jung, "HRas intracellular trafficking and signal transduction" (2007). Retrospective Theses and Dissertations. 13946. https://lib.dr.iastate.edu/rtd/13946 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]. HRas intracellular trafficking and signal transduction by Jodi Ho-Jung McKay A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Genetics Program of Study Committee: Janice E. Buss, Co-major Professor Linda Ambrosio, Co-major Professor Diane Bassham Drena Dobbs Ted Huiatt Iowa State University Ames, Iowa 2007 Copyright © Jodi Ho-Jung McKay, 2007. All rights reserved. UMI Number: 3274881 Copyright 2007 by McKay, Jodi Ho-Jung All rights reserved. UMI Microform 3274881 Copyright 2008 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, MI 48106-1346 ii All men dream but not equally. Those who dream by night in the dusty recesses of their minds wake in the day to find that it was vanity; but the dreamers of the day are dangerous men, for they may act their dreams with open eyes, to make it possible. T. E. Lawrence, Seven Pillars of Wisdom, a Triumph iii Table of Contents List of Figures.......................................................................................................................viii List of Tables ........................................................................................................................... x Abstract................................................................................................................................... xi Chapter 1 General Introduction............................................................................................ 1 Molecular responses .............................................................................................................2 Cellular responses................................................................................................................. 3 Thesis Organization .................................................................................................................. 5 HRas does not need COP I- or COP II-dependent vesicular transport to reach the plasma membrane ..................................................................................................... 5 Golgi–independent trafficking requires the conservation of HRas hypervariable region.............................................................................................................................. 6 Internalization of HRas via Arf6-associated endosomal pathway........................................ 6 Chapter 5 General conclusions ............................................................................................. 7 General Literature Review........................................................................................................ 7 Posttranslational modification .............................................................................................. 8 Ras signal transduction .........................................................................................................9 Ras mediated signal transduction from endomembranes ................................................... 10 Vesicular endocytosis ......................................................................................................... 11 Phagocytosis ................................................................................................................... 12 Pinocytosis ...................................................................................................................... 12 Endosomal Recycling Center.......................................................................................... 13 Vesicular exocytosis ........................................................................................................... 14 Cargo transport from ER to Golgi .................................................................................. 14 Cargo transport from Golgi to ER .................................................................................. 15 Microtubules and intracellular trafficking .......................................................................... 16 Concluding Remarks............................................................................................................... 17 References............................................................................................................................... 17 Figure Legends........................................................................................................................ 28 iv Figures..................................................................................................................................... 31 H-Ras does not need COP I- or COP II-dependent vesicular transport to reach the plasma membrane.............................................................................................................. 37 Abstract................................................................................................................................... 37 Introduction............................................................................................................................. 38 Materials and Methods............................................................................................................ 41 Mutants, Plasmids and Antibodies...................................................................................... 41 Cell Culture and transfection .............................................................................................. 41 Counting cells ..................................................................................................................... 41 Brefeldin A, nocodazole and cycloheximide treatment...................................................... 42 Immuno-fluorescence imaging ........................................................................................... 42 Movies ................................................................................................................................ 43 Results..................................................................................................................................... 43 H-Ras proteins often show little accumulation in the Golgi............................................... 43 Nascent H-Ras moves much faster than YFP-GT46 to the plasma membrane .................. 45 Neither Brefeldin A nor dominant negative Sar1 prevent H-Ras movement to the plasma membrane...................................................................................................................... 47 Structures that carry H-Ras do not show directed movement to the plasma membrane in BFA-treated cells .......................................................................................................... 50 Nocodazole does not prevent H-Ras movement to the plasma membrane......................... 52 Some H-Ras accumulates at the ERGIC when the classical pathway is blocked............... 53 H-Ras that is in the ERGIC does not access the alternate pathway in nocodazole-treated cells ............................................................................................................................... 54 Discussion............................................................................................................................... 56 Acknowledgements................................................................................................................. 59 References............................................................................................................................... 59 Figure Legends........................................................................................................................ 61 Table Legends......................................................................................................................... 64 Figures..................................................................................................................................... 66 Trafficking requires conservation of the HRas hypervariable region............................. 74 v Abstract................................................................................................................................... 74 Introduction............................................................................................................................
Load more

Recommended publications
  • REVIEW Signal Transduction, Cell Cycle Regulatory, and Anti
    Leukemia (1999) 13, 1109–1166 1999 Stockton Press All rights reserved 0887-6924/99 $12.00 http://www.stockton-press.co.uk/leu REVIEW Signal transduction, cell cycle regulatory, and anti-apoptotic pathways regulated by IL-3 in hematopoietic cells: possible sites for intervention with anti-neoplastic drugs WL Blalock1, C Weinstein-Oppenheimer1,2, F Chang1, PE Hoyle1, X-Y Wang3, PA Algate4, RA Franklin1,5, SM Oberhaus1,5, LS Steelman1 and JA McCubrey1,5 1Department of Microbiology and Immunology, 5Leo Jenkins Cancer Center, East Carolina University School of Medicine Greenville, NC, USA; 2Escuela de Quı´mica y Farmacia, Facultad de Medicina, Universidad de Valparaiso, Valparaiso, Chile; 3Department of Laboratory Medicine and Pathology, Mayo Clinic and Foundation, Rochester, MN, USA; and 4Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, USA Over the past decade, there has been an exponential increase growth factor), Flt-L (the ligand for the flt2/3 receptor), erythro- in our knowledge of how cytokines regulate signal transduc- poietin (EPO), and others affect the growth and differentiation tion, cell cycle progression, differentiation and apoptosis. Research has focused on different biochemical and genetic of these early hematopoietic precursor cells into cells of the 1–4 aspects of these processes. Initially, cytokines were identified myeloid, lymphoid and erythroid lineages (Table 1). This by clonogenic assays and purified by biochemical techniques. review will concentrate on IL-3 since much of the knowledge This soon led to the molecular cloning of the genes encoding of how cytokines affect cell growth, signal transduction, and the cytokines and their cognate receptors.
    [Show full text]
  • Myopia Genetics Report
    Special Issue IMI – Myopia Genetics Report Milly S. Tedja,1,2 Annechien E. G. Haarman,1,2 Magda A. Meester-Smoor,1,2 Jaakko Kaprio,3,4 David A. Mackey,5–7 Jeremy A. Guggenheim,8 Christopher J. Hammond,9 Virginie J. M. Verhoeven,1,2,10 and Caroline C. W. Klaver1,2,11; for the CREAM Consortium 1Department of Ophthalmology, Erasmus Medical Center, Rotterdam, the Netherlands 2Department of Epidemiology, Erasmus Medical Center, Rotterdam, the Netherlands 3Institute for Molecular Medicine, University of Helsinki, Helsinki, Finland 4Department of Public Health, University of Helsinki, Helsinki, Finland 5Centre for Eye Research Australia, Ophthalmology, Department of Surgery, University of Melbourne, Royal Victorian Eye and Ear Hospital, Melbourne, Victoria, Australia 6Department of Ophthalmology, Menzies Institute of Medical Research, University of Tasmania, Hobart, Tasmania, Australia 7Centre for Ophthalmology and Visual Science, Lions Eye Institute, University of Western Australia, Perth, Western Australia, Australia 8School of Optometry and Vision Sciences, Cardiff University, Cardiff, United Kingdom 9Section of Academic Ophthalmology, School of Life Course Sciences, King’s College London, London, United Kingdom 10Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, the Netherlands 11Department of Ophthalmology, Radboud University Medical Center, Nijmegen, the Netherlands Correspondence: Caroline C. W. The knowledge on the genetic background of refractive error and myopia has expanded Klaver, Erasmus Medical Center, dramatically in the past few years. This white paper aims to provide a concise summary of Room Na-2808, P.O. Box 2040, 3000 current genetic findings and defines the direction where development is needed. CA, Rotterdam, the Netherlands; [email protected]. We performed an extensive literature search and conducted informal discussions with key MST and AEGH contributed equally to stakeholders.
    [Show full text]
  • ADP-Ribosylation Factor, a Small GTP-Binding Protein, Is Required for Binding of the Coatomer Protein Fl-COP to Golgi Membranes JULIE G
    Proc. Natl. Acad. Sci. USA Vol. 89, pp. 6408-6412, July 1992 Biochemistry ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein fl-COP to Golgi membranes JULIE G. DONALDSON*, DAN CASSEL*t, RICHARD A. KAHN*, AND RICHARD D. KLAUSNER* *Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, and tLaboratory of Biological Chemistry, Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 Communicated by Marc Kirschner, April 20, 1992 (receivedfor review February 11, 1992) ABSTRACT The coatomer is a cytosolic protein complex localized to the Golgi complex, although their functions have that reversibly associates with Golgi membranes and is Impli- not been defined. Distinct among these proteins is the ADP- cated in modulating Golgi membrane transport. The associa- ribosylation factor (ARF), originally identified as a cofactor tion of 13-COP, a component of coatomer, with Golgi mem- required for in vitro cholera toxin-catalyzed ADP- branes is enhanced by guanosine 5'-[v-thioltriphosphate ribosylation of the a subunit of the trimeric GTP-binding (GTP[yS]), a nonhydrolyzable analogue of GTP, and by a protein G, (G,.) (19). ARF is an abundant cytosolic protein mixture of aluminum and fluoride ions (Al/F). Here we show that reversibly associates with Golgi membranes (20, 21). that the ADP-ribosylation factor (ARF) is required for the ARF has been shown to be present on Golgi coated vesicles binding of (-COP. Thus, 13-COP contained in a coatomer generated in the presence of GTP[yS], but it is not a com- fraction that has been resolved from ARF does not bind to Golgi ponent of the cytosolic coatomer (22).
    [Show full text]
  • G-Protein ␤␥-Complex Is Crucial for Efficient Signal Amplification in Vision
    The Journal of Neuroscience, June 1, 2011 • 31(22):8067–8077 • 8067 Cellular/Molecular G-Protein ␤␥-Complex Is Crucial for Efficient Signal Amplification in Vision Alexander V. Kolesnikov,1 Loryn Rikimaru,2 Anne K. Hennig,1 Peter D. Lukasiewicz,1 Steven J. Fliesler,4,5,6,7 Victor I. Govardovskii,8 Vladimir J. Kefalov,1 and Oleg G. Kisselev2,3 1Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri 63110, Departments of 2Ophthalmology and 3Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Saint Louis, Missouri 63104, 4Research Service, Veterans Administration Western New York Healthcare System, and Departments of 5Ophthalmology (Ross Eye Institute) and 6Biochemistry, University at Buffalo/The State University of New York (SUNY), and 7SUNY Eye Institute, Buffalo, New York 14215, and 8Sechenov Institute for Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Saint Petersburg 194223, Russia A fundamental question of cell signaling biology is how faint external signals produce robust physiological responses. One universal mechanism relies on signal amplification via intracellular cascades mediated by heterotrimeric G-proteins. This high amplification system allows retinal rod photoreceptors to detect single photons of light. Although much is now known about the role of the ␣-subunit of the rod-specific G-protein transducin in phototransduction, the physiological function of the auxiliary ␤␥-complex in this process remains a mystery. Here, we show that elimination of the transducin ␥-subunit drastically reduces signal amplification in intact mouse rods. The consequence is a striking decline in rod visual sensitivity and severe impairment of nocturnal vision. Our findings demonstrate that transducin ␤␥-complex controls signal amplification of the rod phototransduction cascade and is critical for the ability of rod photoreceptors to function in low light conditions.
    [Show full text]
  • Brief Definitive Report ZAP- 70 Gene
    Brief Definitive Report Reconstitution of T Cell Receptor Signaling in ZAP-70-deficient Cells by Retroviral Transduction of the ZAP- 70 Gene By Naomi Taylor,* Kevin B. Bacon,¢ Susan Smith,* Thomas Jahn,* Theresa A. Kadlecekfl Lisa Uribe,* Donald B. Kohn,* Erwin W. Gelfand,IIArthur Weiss,~ and Kenneth Weinberg* From the *Division of Research Immunology and Bone Marrow Transplantation, Children's Hospital Los Angeles, Los Angeles, California 90027; :~DNAX Research Institute, Palo Alto, California 94304; gHoward Hughes Medical Institute, Department of Medicine and of Microbiology and Immunology, University of California, San Francisco, California 94143; and IIDivision of Basic Sciences and Molecular Signal Transduction Program, Department of Pediatrics, National Jewish Centerfor Immunology and Respiratory Diseases, Denver, Colorado 80206 Summal-y A variant of severe combined lmmunodeficiency syndrome (SCID) with a selective inability to produce CD8 single positive T cells and a signal transduction defect in peripheral CD4 + cells has recently been shown to be the result of mutations in the ZAP-70 gene. T cell receptor (TCR) signaling requires the association of the ZAP-70 protein tyrosine kinase with the TCR complex. Human T cell leukemia virus type I-transformed CD4 + T cell lines w.ere established from ZAP-70-deficient patients and normal controls. ZAP-70 was expressed and appropriately phosphorylated in normal T cell lines after TCR engagement, but was not detected in T cell lines from ZAP-70-deficient patients. To determine whether signaling could be reconstituted, wild-type ZAP-70 was introduced into deficient cells with a ZAP-70 retroviral vector. High titer producer clones expressing ZAP-70 were generated in the Gibbon ape leukemia virus packaging line PG13.
    [Show full text]
  • Identification of HRAS Mutations and Absence of GNAQ Or GNA11
    Modern Pathology (2013) 26, 1320–1328 1320 & 2013 USCAP, Inc All rights reserved 0893-3952/13 $32.00 Identification of HRAS mutations and absence of GNAQ or GNA11 mutations in deep penetrating nevi Ryan P Bender1, Matthew J McGinniss2, Paula Esmay1, Elsa F Velazquez3,4 and Julie DR Reimann3,4 1Caris Life Sciences, Phoenix, AZ, USA; 2Genoptix Medical Laboratory, Carlsbad, CA, USA; 3Dermatopathology Division, Miraca Life Sciences Research Institute, Newton, MA, USA and 4Department of Dermatology, Tufts Medical Center, Boston, MA, USA HRAS is mutated in B15% of Spitz nevi, and GNAQ or GNA11 is mutated in blue nevi (46–83% and B7% respectively). Epithelioid blue nevi and deep penetrating nevi show features of both blue nevi (intradermal location, pigmentation) and Spitz nevi (epithelioid morphology). Epithelioid blue nevi and deep penetrating nevi can also show overlapping features with melanoma, posing a diagnostic challenge. Although epithelioid blue nevi are considered blue nevic variants, no GNAQ or GNA11 mutations have been reported. Classification of deep penetrating nevi as blue nevic variants has also been proposed, however, no GNAQ or GNA11 mutations have been reported and none have been tested for HRAS mutations. To better characterize these tumors, we performed mutational analysis for GNAQ, GNA11, and HRAS, with blue nevi and Spitz nevi as controls. Within deep penetrating nevi, none demonstrated GNAQ or GNA11 mutations (0/38). However, 6% revealed HRAS mutation (2/32). Twenty percent of epithelioid blue nevi contained a GNAQ mutation (2/10), while none displayed GNA11 or HRAS mutation. Eighty-seven percent of blue nevi contained a GNAQ mutation (26/30), 4% a GNA11 mutation (1/28), and none an HRAS mutation.
    [Show full text]
  • Prevalence of Mutations in TSHR, GNAS, PRKAR1A and RAS Genes
    European Journal of Endocrinology (2008) 159 623–631 ISSN 0804-4643 CLINICAL STUDY Prevalence of mutations in TSHR, GNAS, PRKAR1A and RAS genes in a large series of toxic thyroid adenomas from Galicia, an iodine-deficient area in NW Spain F Palos-Paz1, O Perez-Guerra1, J Cameselle-Teijeiro3,CRueda-Chimeno5, F Barreiro-Morandeira4, J Lado-Abeal1,2 and the Galician Group for the Study of Toxic Multinodular Goitre: D Araujo Vilar1,2, R Argueso7, O Barca1, MBotana7, J M Cabezas-Agrı´cola2, P Catalina6, L Dominguez Gerpe1, T Fernandez9, A Mato8, A Nun˜o11,MPenin10 and B Victoria1 1Unidade de Enfermedades Tiroideas e Metabo´licas (UETeM), 2Endocrinology Section, Department of Medicine, 3Pathology Department and 4Surgery Department, Complexo Hospitalario Universitary de Santiago (CHUS), University of Santiago de Compostela, Santiago de Compostela, 15705, Spain, 5General Surgery Section and 6Endocrinology Section, Complexo Hospitalario de Pontevedra, Pontevedra, Spain, 7Endocrinology Section, Complexo Hospitalario Xeral-Calde, Lugo, Spain, 8Endocrinology Section, Complexo Hospitalario de Ourense, Ourense, Spain, 9Endocrinology Section, Complexo Hospitalario Universitario Juan-Canalejo, A Corun˜a, Spain, 10Endocrinology Section, Hospital Arquitecto Marcide, Ferrol, Spain and 11General Surgery Section, Hospital do Meixoeiro, Complexo Hospitalario Universitario de Vigo, Vigo, Spain (Correspondence should be addressed to J Lado-Abeal who is now at UETeM, Department of Medicine, School of Medicine, University of Santiago de Compostela, C/San Francisco sn 15705, Santiago de Compostela, Spain; Email: [email protected]) Abstract Objective: Toxic thyroid adenoma (TA) is a common cause of hyperthyroidism. Mutations in the TSH receptor (TSHR) gene, and less frequently in the adenylate cyclase-stimulating G alpha protein (GNAS) gene, are well established causes of TA in Europe.
    [Show full text]
  • Mosaic Activating Mutations in GNA11 and GNAQ Are Associated with Phakomatosis Pigmentovascularis and Extensive Dermal Melanocytosis Anna C
    ORIGINAL ARTICLE Mosaic Activating Mutations in GNA11 and GNAQ Are Associated with Phakomatosis Pigmentovascularis and Extensive Dermal Melanocytosis Anna C. Thomas1,18, Zhiqiang Zeng2,18, Jean-Baptiste Rivie`re3,18, Ryan O’Shaughnessy4, Lara Al-Olabi1, Judith St.-Onge3, David J. Atherton5,He´le`ne Aubert6, Lorea Bagazgoitia7, Se´bastien Barbarot6, Emmanuelle Bourrat8,9, Christine Chiaverini10, W. Kling Chong11, Yannis Duffourd3, Mary Glover5, Leopold Groesser12, Smail Hadj-Rabia13, Henning Hamm14, Rudolf Happle15, Imran Mushtaq16, Jean-Philippe Lacour10, Regula Waelchli5, Marion Wobser14, Pierre Vabres3,17,19, E. Elizabeth Patton2,19 and Veronica A. Kinsler1,5,19 Common birthmarks can be an indicator of underlying genetic disease but are often overlooked. Mongolian blue spots (dermal melanocytosis) are usually localized and transient, but they can be extensive, permanent, and associated with extracutaneous abnormalities. Co-occurrence with vascular birthmarks defines a subtype of phakomatosis pigmentovascularis, a group of syndromes associated with neurovascular, ophthalmological, overgrowth, and malignant complications. Here, we discover that extensive dermal melanocytosis and pha- komatosis pigmentovascularis are associated with activating mutations in GNA11 and GNAQ, genes that encode Ga subunits of heterotrimeric G proteins. The mutations were detected at very low levels in affected tissues but were undetectable in the blood, indicating that these conditions are postzygotic mosaic disorders. R183C Q209L In vitro expression of mutant GNA11 and GNA11 in human cell lines demonstrated activation of the downstream p38 MAPK signaling pathway and the p38, JNK, and ERK pathways, respectively. Transgenic R183C mosaic zebrafish models expressing mutant GNA11 under promoter mitfa developed extensive dermal melanocytosis recapitulating the human phenotype. Phakomatosis pigmentovascularis and extensive dermal melanocytosis are therefore diagnoses in the group of mosaic heterotrimeric G-protein disorders, joining McCune-Albright and Sturge-Weber syndromes.
    [Show full text]
  • Identification of Motifs in Cholera Toxin A1 Polypeptide That Are Required for Its Interaction with Human ADP-Ribosylation Factor 6 in a Bacterial Two-Hybrid System
    Identification of motifs in cholera toxin A1 polypeptide that are required for its interaction with human ADP-ribosylation factor 6 in a bacterial two-hybrid system Michael G. Jobling and Randall K. Holmes* Department of Microbiology, University of Colorado Health Sciences Center, Denver, CO 80220 Edited by John J. Mekalanos, Harvard Medical School, Boston, MA, and approved October 20, 2000 (received for review September 14, 2000) The latent ADP-ribosyltransferase activity of cholera toxin (CT) that teins called ADP-ribosylation factors (ARFs; refs. 5 and 6). is activated after proteolytic nicking and reduction is associated ARFs are members of a highly conserved multigene family of with the CT A1 subunit (CTA1) polypeptide. This activity is stimu- small GTP-binding proteins, originally identified as activators of lated in vitro by interaction with eukaryotic proteins termed CT. They interact with several other proteins and are ubiqui- ADP-ribosylation factors (ARFs). We analyzed this interaction in a tously involved in membrane trafficking events (7). ARFs must modified bacterial two-hybrid system in which the T18 and T25 be in the GTP-bound form to be active. There are six mammalian fragments of the catalytic domain of Bordetella pertussis adenylate ARFs (five human) that fall into three classes: ARFs 1, 2, and cyclase were fused to CTA1 and human ARF6 polypeptides, respec- 3 (class I), ARFs 4 and 5 (class II), and ARF6 (class III). Class tively. Direct interaction between the CTA1 and ARF6 domains in I ARFs regulate the assembly of several types of vesicle coat these hybrid proteins reconstituted the adenylate cyclase activity complexes (8).
    [Show full text]
  • Multi-Functionality of Proteins Involved in GPCR and G Protein Signaling: Making Sense of Structure–Function Continuum with In
    Cellular and Molecular Life Sciences (2019) 76:4461–4492 https://doi.org/10.1007/s00018-019-03276-1 Cellular andMolecular Life Sciences REVIEW Multi‑functionality of proteins involved in GPCR and G protein signaling: making sense of structure–function continuum with intrinsic disorder‑based proteoforms Alexander V. Fonin1 · April L. Darling2 · Irina M. Kuznetsova1 · Konstantin K. Turoverov1,3 · Vladimir N. Uversky2,4 Received: 5 August 2019 / Revised: 5 August 2019 / Accepted: 12 August 2019 / Published online: 19 August 2019 © Springer Nature Switzerland AG 2019 Abstract GPCR–G protein signaling system recognizes a multitude of extracellular ligands and triggers a variety of intracellular signal- ing cascades in response. In humans, this system includes more than 800 various GPCRs and a large set of heterotrimeric G proteins. Complexity of this system goes far beyond a multitude of pair-wise ligand–GPCR and GPCR–G protein interactions. In fact, one GPCR can recognize more than one extracellular signal and interact with more than one G protein. Furthermore, one ligand can activate more than one GPCR, and multiple GPCRs can couple to the same G protein. This defnes an intricate multifunctionality of this important signaling system. Here, we show that the multifunctionality of GPCR–G protein system represents an illustrative example of the protein structure–function continuum, where structures of the involved proteins represent a complex mosaic of diferently folded regions (foldons, non-foldons, unfoldons, semi-foldons, and inducible foldons). The functionality of resulting highly dynamic conformational ensembles is fne-tuned by various post-translational modifcations and alternative splicing, and such ensembles can undergo dramatic changes at interaction with their specifc partners.
    [Show full text]
  • Supplementary Table 2
    Supplementary Table 2. Differentially Expressed Genes following Sham treatment relative to Untreated Controls Fold Change Accession Name Symbol 3 h 12 h NM_013121 CD28 antigen Cd28 12.82 BG665360 FMS-like tyrosine kinase 1 Flt1 9.63 NM_012701 Adrenergic receptor, beta 1 Adrb1 8.24 0.46 U20796 Nuclear receptor subfamily 1, group D, member 2 Nr1d2 7.22 NM_017116 Calpain 2 Capn2 6.41 BE097282 Guanine nucleotide binding protein, alpha 12 Gna12 6.21 NM_053328 Basic helix-loop-helix domain containing, class B2 Bhlhb2 5.79 NM_053831 Guanylate cyclase 2f Gucy2f 5.71 AW251703 Tumor necrosis factor receptor superfamily, member 12a Tnfrsf12a 5.57 NM_021691 Twist homolog 2 (Drosophila) Twist2 5.42 NM_133550 Fc receptor, IgE, low affinity II, alpha polypeptide Fcer2a 4.93 NM_031120 Signal sequence receptor, gamma Ssr3 4.84 NM_053544 Secreted frizzled-related protein 4 Sfrp4 4.73 NM_053910 Pleckstrin homology, Sec7 and coiled/coil domains 1 Pscd1 4.69 BE113233 Suppressor of cytokine signaling 2 Socs2 4.68 NM_053949 Potassium voltage-gated channel, subfamily H (eag- Kcnh2 4.60 related), member 2 NM_017305 Glutamate cysteine ligase, modifier subunit Gclm 4.59 NM_017309 Protein phospatase 3, regulatory subunit B, alpha Ppp3r1 4.54 isoform,type 1 NM_012765 5-hydroxytryptamine (serotonin) receptor 2C Htr2c 4.46 NM_017218 V-erb-b2 erythroblastic leukemia viral oncogene homolog Erbb3 4.42 3 (avian) AW918369 Zinc finger protein 191 Zfp191 4.38 NM_031034 Guanine nucleotide binding protein, alpha 12 Gna12 4.38 NM_017020 Interleukin 6 receptor Il6r 4.37 AJ002942
    [Show full text]
  • Lysophosphatidic Acid Activates Arf6 to Promote the Mesenchymal Malignancy of Renal Cancer
    ARTICLE Received 6 Jun 2015 | Accepted 6 Jan 2016 | Published 8 Feb 2016 DOI: 10.1038/ncomms10656 OPEN Lysophosphatidic acid activates Arf6 to promote the mesenchymal malignancy of renal cancer Shigeru Hashimoto1,*, Shuji Mikami2,*,w, Hirokazu Sugino1, Ayumu Yoshikawa1, Ari Hashimoto1, Yasuhito Onodera1, Shotaro Furukawa1,3, Haruka Handa1, Tsukasa Oikawa1, Yasunori Okada4, Mototsugu Oya5 & Hisataka Sabe1 Acquisition of mesenchymal properties by cancer cells is critical for their malignant behaviour, but regulators of the mesenchymal molecular machinery and how it is activated remain elusive. Here we show that clear cell renal cell carcinomas (ccRCCs) frequently utilize the Arf6-based mesenchymal pathway to promote invasion and metastasis, similar to breast cancers. In breast cancer cells, ligand-activated receptor tyrosine kinases employ GEP100 to activate Arf6, which then recruits AMAP1; and AMAP1 then binds to the mesenchymal- specific protein EPB41L5, which promotes epithelial–mesenchymal transition and focal adhesion dynamics. In renal cancer cells, lysophosphatidic acid (LPA) activates Arf6 via its G-protein-coupled receptors, in which GTP-Ga12 binds to EFA6. The Arf6-based pathway may also contribute to drug resistance. Our results identify a specific mesenchymal molecular machinery of primary ccRCCs, which is triggered by a product of autotaxin and it is associated with poor outcome of patients. 1 Department of Molecular Biology, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan. 2 Division of Diagnostic Pathology, Keio University Hospital, Tokyo 160-0016, Japan. 3 Department of Gastroenterological Surgery II, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan. 4 Department of Pathology, Keio University School of Medicine, Tokyo 160-0016, Japan.
    [Show full text]