DOCSLIB.ORG

Phosphatidylinositol 3-Kinase Kinase-3 Signaling Pathway Via Akt

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Phosphatidylinositol 3-Kinase Kinase-3 Signaling Pathway Via Akt The B Cell Antigen Receptor Activates the Akt (Protein Kinase B)/Glycogen Synthase Kinase-3 Signaling Pathway Via Phosphatidylinositol 3-Kinase This information is current as of September 28, 2021. Michael R. Gold, Michael P. Scheid, Lorna Santos, May Dang-Lawson, Richard A. Roth, Linda Matsuuchi, Vincent Duronio and Danielle L. Krebs J Immunol 1999; 163:1894-1905; ; http://www.jimmunol.org/content/163/4/1894 Downloaded from References This article cites 82 articles, 45 of which you can access for free at: http://www.jimmunol.org/content/163/4/1894.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 28, 2021 *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 © 1999 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The B Cell Antigen Receptor Activates the Akt (Protein Kinase B)/Glycogen Synthase Kinase-3 Signaling Pathway Via Phosphatidylinositol 3-Kinase1 Michael R. Gold,2* Michael P. Scheid,† Lorna Santos,‡ May Dang-Lawson,* Richard A. Roth,§ Linda Matsuuchi,‡ Vincent Duronio,† and Danielle L. Krebs3* We have previously shown that the B cell Ag receptor (BCR) activates phosphatidylinositol (PI) 3-kinase. We now show that a serine/threonine kinase called Akt or protein kinase B is a downstream target of PI 3-kinase in B cells. Akt has been shown to promote cell survival as well as the transcription and translation of proteins involved in cell cycle progression. Using an Ab that specifically recognizes the activated form of Akt that is phosphorylated on serine 473, we show that BCR engagement activates Akt in a PI 3-kinase-dependent manner. These results were confirmed using in vitro kinase assays. Moreover, BCR ligation also Downloaded from induced phosphorylation of Akt of threonine 308, another modification that is required for activation of Akt. In the DT40 chicken B cell line, phosphorylation of Akt on serine 473 was completely dependent on the Lyn tyrosine kinase, while the Syk tyrosine kinase was required for sustained phosphorylation of Akt. Complementary experiments in BCR-expressing AtT20 endocrine cells confirmed that Src kinases are sufficient for BCR-induced Akt phosphorylation, but that Syk is required for sustained phosphor- ylation of Akt on both serine 473 and threonine 308. In insulin-responsive cells, Akt phosphorylates and inactivates the serine/ threonine kinase glycogen synthase kinase-3 (GSK-3). Inactivation of GSK-3 may promote nuclear accumulation of several http://www.jimmunol.org/ transcription factors, including NF-ATc. We found that BCR engagement induced GSK-3 phosphorylation and decreased GSK-3 enzyme activity. Thus, BCR ligation initiates a PI 3-kinase/Akt/GSK-3 signaling pathway. The Journal of Immunology, 1999, 163: 1894–1905. ngagement of the B cell Ag receptor (BCR)4 by Ags ini- line (4, 5), until recently little was known about the downstream tiates signals that can promote apoptosis, anergy, survival, targets of PI 3-kinase signaling in B cells. E activation, or proliferation, depending upon the differen- Recent work has revealed that PI 3-kinase plays a key role in tiation state of the B cell, the magnitude, and duration of the BCR multiple cellular processes. PI 3,4,5-trisphosphate (PIP3), a plasma by guest on September 28, 2021 signal, and whether or not the B cell receives additional signals membrane phospholipid produced by PI 3-kinase, is a ligand for from CD40 or cytokine receptors (1, 2). When clustered by mul- the pleckstrin homology (PH) domains that are found in a variety tivalent Ags or by anti-Ig Abs, the BCR activates three key sig- of cytosolic proteins, many of which are involved in signal trans- naling enzymes: Ras, phospholipase C-g (PLC-g), and phosphati- duction (6). By binding PH domains, PIP3 can recruit PH domain- dylinositol (PI) 3-kinase (1, 3). Although PI 3-kinase had been containing proteins to the plasma membrane. This is of particular shown to contribute to the ability of the BCR to regulate the pro- importance for cytosolic signaling enzymes that act on membrane- liferation of normal B cells, as well as a human B lymphoma cell associated substrates. In addition to recruiting signaling proteins to the plasma membrane, the binding of PIP3 to a protein’s PH do- main may induce conformational changes that affect its enzymatic Departments of *Microbiology and Immunology, †Medicine, and ‡Zoology, Univer- activity, its ability to act as a substrate for kinases, or its ability to sity of British Columbia, Vancouver, British Columbia, Canada; and §Department of interact with other proteins. Molecular Pharmacology, Stanford University School of Medicine, Stanford, CA In B cells, PIP -PH domain interactions play an essential role in 94305 3 the ability of the BCR to stimulate PLC-g-dependent signaling. Received for publication February 18, 1999. Accepted for publication June 9, 1999. The Btk tyrosine kinase, which phosphorylates and activates The costs of publication of this article were defrayed in part by the payment of page g charges. This article must therefore be hereby marked advertisement in accordance PLC- , is recruited to the plasma membrane via the binding of its with 18 U.S.C. Section 1734 solely to indicate this fact. PH domain to PIP3 (7–10). PIP3-PH domain interactions are also 1 This work was supported by grants from the Medical Research Council of Canada involved in activation of the Rac1 GTPase, which is required for (to M.R.G., L.M., and V.D), a grant from the Natural Sciences and Engineering sustained hydrolysis of PI 4,5-bisphosphate by PLC-g (11). Rac1- Research Council of Canada (to M.R.G.), Medical Research Council Scholarships (to M.R.G. and V.D.), a Cancer Research Society (Canada) studentship (to M.P.S.), and GTP activates PI 4-phosphate 5-kinases, which are responsible for a University of British Columbia Graduate Fellowship (to D.L.K.). providing a continuous supply of PI 4,5-bisphosphate to act as a 2 Address correspondence and reprint requests to Dr. Michael R. Gold, Department of substrate for PLC-g (12–14). Both Sos and Vav can act as ex- Microbiology and Immunology, University of British Columbia, 6174 University Boulevard, Vancouver, British Columbia, Canada V6T 1Z3. E-mail address: change factors that activate Rac1, and their ability to do so is [email protected] dependent on the binding of PIP3 to their PH domains (15, 16). In 3 Current address: Walter and Eliza Hall Institute for Medical Research, Melbourne, addition to Btk, SOS, and Vav, a large number of other PH domain- Victoria 3050, Australia. containing proteins have been identified, many of them cytosolic sig- 4 Abbreviations used in this paper: BCR, B cell Ag receptor; PLC-g, phospholipase naling proteins (6). Thus, it is likely that the activation of PI 3-kinase g C- , PI, phosphatidylinositol; PIP3, PI 3,4,5-trisphosphate; PH, pleckstrin homology; by the BCR and the subsequent production of PIP3 regulates many PI(3,4)P2, PI 3,4-bisphosphate; ILK, integrin-linked kinase; GSK-3, glycogen syn- thase kinase-3; HA, hemagglutinin. other signaling pathways in addition to the PLC-g pathway. Copyright © 1999 by The American Association of Immunologists 0022-1767/99/$02.00 The Journal of Immunology 1895 There is now considerable evidence that a 60-kDa serine/thre- forms of Akt also selectively enhance the translation of cyclin D1 onine protein kinase called Akt (or protein kinase B) is a major (47). Thus, Akt may promote cell cycle progression at both the downstream target of PI 3-kinase signaling. Akt is activated by a transcriptional and posttranscriptional levels. number of receptors that activate PI 3-kinase, including the recep- Another major downstream target of Akt is glycogen synthase tors for insulin, platelet-derived growth factor, epidermal growth kinase-3 (GSK-3), a constitutively active serine/threonine kinase factor, IL-2, IL-3, IL-4, GM-CSF, and stem cell factor (17–24). In whose activity is inhibited by Akt. Regulation of GSK-3 activity these systems, it has been clearly shown that activation of PI 3-ki- has been studied primarily in the context of insulin receptor sig- nase is both necessary and sufficient for Akt to be activated. Re- naling. Insulin receptor signaling leads to a reduction in GSK-3 ceptor-induced activation of Akt is blocked by PI 3-kinase inhib- activity (20, 48, 49), and this appears to be mediated by Akt, since itors and by expression of dominant negative forms of PI 3-kinase a dominant-negative form of Akt can block the ability of insulin to (17–24). Conversely, expression of constitutively active forms of decrease GSK-3 activity (48). In vitro, Akt can phosphorylate PI 3-kinase in cells results in activation of Akt (25). GSK-3a on serine 21 and GSK-3b on serine 9, and these modi- Considerable progress has been made toward understanding fications correlate with inhibition of GSK-3 enzyme activity (20). how PI 3-kinase activates Akt (reviewed in Ref. 26). Akt has a PH Thus, GSK-3a and GSK-3b may be physiological substrates domain that binds the PI 3-kinase-derived lipids PIP and PI 3,4- 3 of Akt.
Load more

Recommended publications
  • Supplemental Information to Mammadova-Bach Et Al., “Laminin Α1 Orchestrates VEGFA Functions in the Ecosystem of Colorectal Carcinogenesis”
    Supplemental information to Mammadova-Bach et al., “Laminin α1 orchestrates VEGFA functions in the ecosystem of colorectal carcinogenesis” Supplemental material and methods Cloning of the villin-LMα1 vector The plasmid pBS-villin-promoter containing the 3.5 Kb of the murine villin promoter, the first non coding exon, 5.5 kb of the first intron and 15 nucleotides of the second villin exon, was generated by S. Robine (Institut Curie, Paris, France). The EcoRI site in the multi cloning site was destroyed by fill in ligation with T4 polymerase according to the manufacturer`s instructions (New England Biolabs, Ozyme, Saint Quentin en Yvelines, France). Site directed mutagenesis (GeneEditor in vitro Site-Directed Mutagenesis system, Promega, Charbonnières-les-Bains, France) was then used to introduce a BsiWI site before the start codon of the villin coding sequence using the 5’ phosphorylated primer: 5’CCTTCTCCTCTAGGCTCGCGTACGATGACGTCGGACTTGCGG3’. A double strand annealed oligonucleotide, 5’GGCCGGACGCGTGAATTCGTCGACGC3’ and 5’GGCCGCGTCGACGAATTCACGC GTCC3’ containing restriction site for MluI, EcoRI and SalI were inserted in the NotI site (present in the multi cloning site), generating the plasmid pBS-villin-promoter-MES. The SV40 polyA region of the pEGFP plasmid (Clontech, Ozyme, Saint Quentin Yvelines, France) was amplified by PCR using primers 5’GGCGCCTCTAGATCATAATCAGCCATA3’ and 5’GGCGCCCTTAAGATACATTGATGAGTT3’ before subcloning into the pGEMTeasy vector (Promega, Charbonnières-les-Bains, France). After EcoRI digestion, the SV40 polyA fragment was purified with the NucleoSpin Extract II kit (Machery-Nagel, Hoerdt, France) and then subcloned into the EcoRI site of the plasmid pBS-villin-promoter-MES. Site directed mutagenesis was used to introduce a BsiWI site (5’ phosphorylated AGCGCAGGGAGCGGCGGCCGTACGATGCGCGGCAGCGGCACG3’) before the initiation codon and a MluI site (5’ phosphorylated 1 CCCGGGCCTGAGCCCTAAACGCGTGCCAGCCTCTGCCCTTGG3’) after the stop codon in the full length cDNA coding for the mouse LMα1 in the pCIS vector (kindly provided by P.
    [Show full text]
  • ROS Production Induced by BRAF Inhibitor Treatment Rewires
    Cesi et al. Molecular Cancer (2017) 16:102 DOI 10.1186/s12943-017-0667-y RESEARCH Open Access ROS production induced by BRAF inhibitor treatment rewires metabolic processes affecting cell growth of melanoma cells Giulia Cesi, Geoffroy Walbrecq, Andreas Zimmer, Stephanie Kreis*† and Claude Haan† Abstract Background: Most melanoma patients with BRAFV600E positive tumors respond well to a combination of BRAF kinase and MEK inhibitors. However, some patients are intrinsically resistant while the majority of patients eventually develop drug resistance to the treatment. For patients insufficiently responding to BRAF and MEK inhibitors, there is an ongoing need for new treatment targets. Cellular metabolism is such a promising new target line: mutant BRAFV600E has been shown to affect the metabolism. Methods: Time course experiments and a series of western blots were performed in a panel of BRAFV600E and BRAFWT/ NRASmut human melanoma cells, which were incubated with BRAF and MEK1 kinase inhibitors. siRNA approaches were used to investigate the metabolic players involved. Reactive oxygen species (ROS) were measured by confocal microscopy and AZD7545, an inhibitor targeting PDKs (pyruvate dehydrogenase kinase) was tested. Results: We show that inhibition of the RAS/RAF/MEK/ERK pathway induces phosphorylation of the pyruvate dehydrogenase PDH-E1α subunit in BRAFV600E and in BRAFWT/NRASmut harboring cells. Inhibition of BRAF, MEK1 and siRNA knock-down of ERK1/2 mediated phosphorylation of PDH. siRNA-mediated knock-down of all PDKs or the use of DCA (a pan-PDK inhibitor) abolished PDH-E1α phosphorylation. BRAF inhibitor treatment also induced the upregulation of ROS, concomitantly with the induction of PDH phosphorylation.
    [Show full text]
  • PDK1 Acquires PDK2 Activity in the Presence of a Synthetic Peptide
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Research Paper 393 PDK1 acquires PDK2 activity in the presence of a synthetic peptide derived from the carboxyl terminus of PRK2 Anudharan Balendran*†, Antonio Casamayor*†, Maria Deak†, Andrew Paterson†, Piers Gaffney‡, Richard Currie§, C. Peter Downes§ and Dario R. Alessi† Background: Protein kinase B (PKB) is activated by phosphorylation of Thr308 Addresses: *MRC Protein Phosphorylation Unit, and of Ser473. Thr308 is phosphorylated by the 3-phosphoinositide-dependent Department of Biochemistry, University of Dundee, ‡ protein kinase-1 (PDK1) but the identity of the kinase that phosphorylates Dundee DD1 5EH, UK. Ludwig Institute of Cancer Research, London W1P 8BY, UK. §Department of Ser473 (provisionally termed PDK2) is unknown. Biochemistry, University of Dundee, Dundee DD1 5EH, UK. Results: The kinase domain of PDK1 interacts with a region of protein kinase C-related kinase-2 (PRK2), termed the PDK1-interacting fragment (PIF). PIF is Correspondence: Dario R. Alessi E-mail: [email protected] situated carboxy-terminal to the kinase domain of PRK2, and contains a consensus motif for phosphorylation by PDK2 similar to that found in PKBα, †A.B. and A.C. contributed equally to this work. except that the residue equivalent to Ser473 is aspartic acid. Mutation of any of the conserved residues in the PDK2 motif of PIF prevented interaction of PIF Received: 12 January 1999 Revised: 18 February 1999 with PDK1. Remarkably, interaction of PDK1 with PIF, or with a synthetic Accepted: 3 March 1999 peptide encompassing the PDK2 consensus sequence of PIF, converted PDK1 from an enzyme that could phosphorylate only Thr308 of PKBα to one that Published: 8 April 1999 phosphorylates both Thr308 and Ser473 of PKBα in a manner dependent on Current Biology 1999, 9:393–404 phosphatidylinositol (3,4,5) trisphosphate (PtdIns(3,4,5)P3).
    [Show full text]
  • AGC Kinases in Mtor Signaling, in Mike Hall and Fuyuhiko Tamanoi: the Enzymes, Vol
    Provided for non-commercial research and educational use only. Not for reproduction, distribution or commercial use. This chapter was originally published in the book, The Enzymes, Vol .27, published by Elsevier, and the attached copy is provided by Elsevier for the author's benefit and for the benefit of the author's institution, for non-commercial research and educational use including without limitation use in instruction at your institution, sending it to specific colleagues who know you, and providing a copy to your institution’s administrator. All other uses, reproduction and distribution, including without limitation commercial reprints, selling or licensing copies or access, or posting on open internet sites, your personal or institution’s website or repository, are prohibited. For exceptions, permission may be sought for such use through Elsevier's permissions site at: http://www.elsevier.com/locate/permissionusematerial From: ESTELA JACINTO, AGC Kinases in mTOR Signaling, In Mike Hall and Fuyuhiko Tamanoi: The Enzymes, Vol. 27, Burlington: Academic Press, 2010, pp.101-128. ISBN: 978-0-12-381539-2, © Copyright 2010 Elsevier Inc, Academic Press. Author's personal copy 7 AGC Kinases in mTOR Signaling ESTELA JACINTO Department of Physiology and Biophysics UMDNJ-Robert Wood Johnson Medical School, Piscataway New Jersey, USA I. Abstract The mammalian target of rapamycin (mTOR), a protein kinase with homology to lipid kinases, orchestrates cellular responses to growth and stress signals. Various extracellular and intracellular inputs to mTOR are known. mTOR processes these inputs as part of two mTOR protein com- plexes, mTORC1 or mTORC2. Surprisingly, despite the many cellular functions that are linked to mTOR, there are very few direct mTOR substrates identified to date.
    [Show full text]
  • Mtor Signaling in Pulmonary Vascular Disease: Pathogenic Role and Therapeutic Target
    International Journal of Molecular Sciences Review mTOR Signaling in Pulmonary Vascular Disease: Pathogenic Role and Therapeutic Target Aleksandra Babicheva 1, Ayako Makino 2 and Jason X.-J. Yuan 1,* 1 Section of Physiology, Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of California, San Diego, CA 92093, USA; [email protected] 2 Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, 9500 Gilman Drive, MC 0856, La Jolla, CA 92093-0856, USA; [email protected] * Correspondence: [email protected] Abstract: Pulmonary arterial hypertension (PAH) is a progressive and fatal disease without a cure. The exact pathogenic mechanisms of PAH are complex and poorly understood, yet a number of abnormally expressed genes and regulatory pathways contribute to sustained vasoconstriction and vascular remodeling of the distal pulmonary arteries. Mammalian target of rapamycin (mTOR) is one of the major signaling pathways implicated in regulating cell proliferation, migration, differentiation, and protein synthesis. Here we will describe the canonical mTOR pathway, structural and functional differences between mTOR complexes 1 and 2, as well as the crosstalk with other important signaling cascades in the development of PAH. The pathogenic role of mTOR in pulmonary vascular remodel- ing and sustained vasoconstriction due to its contribution to proliferation, migration, phenotypic transition, and gene regulation in pulmonary artery smooth muscle and endothelial cells will be discussed. Despite the progress in our elucidation of the etiology and pathogenesis of PAH over the two last decades, there is a lack of effective therapeutic agents to treat PAH patients representing Citation: Babicheva, A.; Makino, A.; a significant unmet clinical need.
    [Show full text]
  • Akt-Mediated Phosphorylation of CDK2 Regulates Its Dual Role in Cell Cycle Progression and Apoptosis
    Research Article 979 Akt-mediated phosphorylation of CDK2 regulates its dual role in cell cycle progression and apoptosis Subbareddy Maddika1,2,*, Sudharsana Rao Ande1, Emilia Wiechec1,3,4, Lise Lotte Hansen4, Sebastian Wesselborg5,‡ and Marek Los1,2,‡,§,¶ 1Manitoba Institute of Cell Biology, CancerCare Manitoba, and 2Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, R3E 0V9, Canada 3Department of Human Genetics, University of Aarhus, DK-8000 C Aarhus, Denmark 4Department of Experimental and Clinical Radiobiology, Oncology Center, Maria Sklodowka-Curie Memorial Institute, Wybrzeze Armii Krajowej 15, PL-44100 Gliwice, Poland 5Department of Internal Medicine I, University of Tübingen, Tübingen, Germany *Present address: Department of Therapeutic Radiology, Yale School of Medicine, New Haven, CT, USA ‡These authors contributed equally to this work §Present address: BioApplications Enterprises, Winnipeg, Manitoba, Canada ¶Author for correspondence (e-mail: [email protected]) Accepted 6 January 2008 Journal of Cell Science 121, 979-988 Published by The Company of Biologists 2008 doi:10.1242/jcs.009530 Summary Here, we show that CDK2, an S-phase cyclin-dependent kinase, and is defective in cytoplasmic localization, severely affects cell is a novel target for Akt during cell cycle progression and cycle progression at the transition from S to G2-M. Interestingly, apoptosis. Akt phosphorylates CDK2 at threonine 39 residue we also show that the Akt/CDK2 pathway is constitutively both in vitro and in vivo. Although CDK2 threonine 39 activated by some anticancer drugs, such as methotrexate and phosphorylation mediated by Akt enhances cyclin-A binding, docetaxel, and under these conditions it promotes, rather than it is dispensable for its basal binding and the kinase activity.
    [Show full text]
  • A Novel JAK1 Mutant Breast Implant-Associated Anaplastic Large Cell Lymphoma Patient-Derived Xenograft Fostering Pre- Clinical Discoveries
    Cancers 2019 S1 of S18 Supplementary Materials: A Novel JAK1 Mutant Breast Implant-Associated Anaplastic Large Cell Lymphoma Patient-Derived Xenograft Fostering Pre- Clinical Discoveries Danilo Fiore, Luca Vincenzo Cappelli, Paul Zumbo, Jude M. Phillip, Zhaoqi Liu, Shuhua Cheng, Liron Yoffe, Paola Ghione, Federica Di Maggio, Ahmet Dogan, Inna Khodos, Elisa de Stanchina, Joseph Casano, Clarisse Kayembe, Wayne Tam, Doron Betel, Robin Foa’, Leandro Cerchietti, Raul Rabadan, Steven Horwitz, David M. Weinstock and Giorgio Inghirami A B C Figure S1. (A) Histology micrografts on IL89 PDTX show overall similarity between T1 T3 and T7 passages (upper panels). Immunohistochemical stains with the indicated antibodies (anti-CD3, anti- CD25 and anti-CD8 [x20]) (lower panels). (B) Flow cytometry panel comprehensive of the most represented surface T-cell lymphoma markers, including: CD2, CD3, CD4, CD5, CD8, CD16, CD25, CD30, CD56, TCRab, TCRgd. IL89 PDTX passage T3 is here depicted for illustration purposes. (C) Analysis of the TCR gamma specific rearrangement clonality in IL89 diagnostic sample and correspondent PDTX after 1 and 5 passages (T1 and T5). A WT Primary p.G1097D IL89 T1 p.G1097D IL89 T5 p.G1097D IL89 cell line B Figure S2. (A) Sanger sequencing confirms the presence of the JAK1 p.G1097D mutation in IL89 PDTX samples and in the cell line, but the mutation is undetectable in the primary due to the low sensitivity of the technique. (B) Manual backtracking of mutations in the primary tumor using deep sequencing data allowed for the identification of several hits at a very low VAF compared to the PDTX-T5. A B IL89 CTRL 30 CTRL Ruxoli?nib S 20 M Ruxoli?nib A R G 10 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 1 1 1 1 1 WEEKS AFTER ENGRAFTMENT Figure S3.
    [Show full text]
  • Upregulation of SLC2A3 Gene and Prognosis in Colorectal Carcinoma
    Kim et al. BMC Cancer (2019) 19:302 https://doi.org/10.1186/s12885-019-5475-x RESEARCH ARTICLE Open Access Upregulation of SLC2A3 gene and prognosis in colorectal carcinoma: analysis of TCGA data Eunyoung Kim1†, Sohee Jung2†, Won Seo Park3, Joon-Hyop Lee4, Rumi Shin5, Seung Chul Heo5, Eun Kyung Choe6, Jae Hyun Lee7, Kwangsoo Kim2* and Young Jun Chai5* Abstract Background: Upregulation of SLC2A genes that encode glucose transporter (GLUT) protein is associated with poor prognosis in many cancers. In colorectal cancer, studies reporting the association between overexpression of GLUT and poor clinical outcomes were flawed by small sample sizes or subjective interpretation of immunohistochemical staining. Here, we analyzed mRNA expressions in all 14 SLC2A genes and evaluated the association with prognosis in colorectal cancer using data from the Cancer Genome Atlas (TCGA) database. Methods: In the present study, we analyzed the expression of SLC2A genes in colorectal cancer and their association with prognosis using data obtained from the TCGA for the discovery sample, and a dataset from the Gene Expression Omnibus for the validation sample. Results: SLC2A3 was significantly associated with overall survival (OS) and disease-free survival (DFS) in both the discovery sample (345 patients) and validation sample (501 patients). High SLC2A3 expression resulted in shorter OS and DFS. In multivariate analyses, high SLC2A3 levels predicted unfavorable OS (adjusted HR 1.95, 95% CI 1.22–3.11; P = 0.005) and were associated with poor DFS (adjusted HR 1.85, 95% CI 1.10–3.12; P = 0.02). Similar results were found in the discovery set.
    [Show full text]
  • Synergistic Inhibitory Effects of Hypoxia and Iron Deficiency On
    Diabetes Volume 65, June 2016 1521 Hyeyoung Nam,1 Deborah Jones,1 Robert C. Cooksey,1 Yan Gao,2 Sandy Sink,2 James Cox,3 and Donald A. McClain1,2,3 Synergistic Inhibitory Effects of Hypoxia and Iron Deficiency on Hepatic Glucose Response in Mouse Liver Diabetes 2016;65:1521–1533 | DOI: 10.2337/db15-0580 Hypoxia and iron both regulate metabolism through mul- Hypoxia results in stabilization of hypoxia-inducible factors tiple mechanisms, including hypoxia-inducible transcrip- (HIFs), transcription factors that control a wide range of tion factors. The hypoxic effects on glucose disposal physiologic processes. HIF-1 and -2 form heterodimeric and glycolysis are well established, but less is known DNA binding complexes composed of a-andb-subunits about the effects of hypoxia and iron deficiency on (3). HIF-1a and HIF-2a are highly structurally homolo- hepatic gluconeogenesis. We therefore assessed their gous; however, the phenotypes of HIF-1a and HIF-2a effects on hepatic glucose production in mice. Weanling knockout mice differ in the effects on transcriptional tar- C57BL/6 male mice were fed an iron-deficient (4 ppm) gets and their tissue distribution (4). Under normoxia, METABOLISM or iron-adequate (35 ppm) diet for 14 weeks and were HIF-a subunits are hydroxylated at proline residues by continued in normoxia or exposed to hypoxia (8% O ) 2 prolyl hydroxylase (PHD) and then rapidly degraded by for the last 4 weeks of that period. Hypoxic mice be- the ubiquitin-proteasome pathway. PHDs are 2-oxoglutarate– came hypoglycemic and displayed impaired hepatic dependent dioxygenases that use O2 as a substrate and iron glucose production after a pyruvate challenge, an ef- fi fect accentuated by an iron-deficient diet.
    [Show full text]
  • Novel Regulation of Mtor Complex 1 Signaling by Site-Specific Mtor Phosphorylation
    Novel Regulation of mTOR Complex 1 Signaling by Site-Specific mTOR Phosphorylation by Bilgen Ekim Üstünel A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Cell and Developmental Biology) in The University of Michigan 2012 Doctoral Committee: Assistant Professor Diane C. Fingar, Chair Associate Professor Billy Tsai Associate Professor Anne B. Vojtek Assistant Professor Patrick J. Hu Assistant Professor Ken Inoki “Our true mentor in life is science.” (“Hayatta en hakiki mürşit ilimdir.”) Mustafa Kemal Atatürk, the founder of Turkish Republic © Bilgen Ekim Üstünel 2012 Acknowledgements This thesis would not have been possible without the enormous support and encouragement of my Ph.D. advisor Diane C. Fingar. I am sincerely thankful for her research insight and guidance during my Ph.D. training. I would like to express my great appreciation to Billy Tsai, Anne B. Vojtek, Ken Inoki, and Patrick J. Hu for serving on my thesis committee, whose advice and help have been valuable. I would like to thank all members of the Fingar, Tsai, and Verhey labs for the discussion in our group meetings. I also would like to thank the CDB administrative staff, especillay Kristen Hug, for their help. I thank Ed Feener for performing the liquid chromatography tandem mass spectrometry analysis to identify novel phosphorylation sites on mTOR and Steve Riddle for performing the in vitro kinome screen to identify candidate kinases for mTOR S2159 phosphorylation site. I thank Brian Magnuson, Hugo A. Acosta-Jaquez, and Jennifer A. Keller for contributing to my first-author paper published in Molecular and Cellular Biology Journal in 2011.
    [Show full text]
  • PRODUCTS and SERVICES Target List
    PRODUCTS AND SERVICES Target list Kinase Products P.1-11 Kinase Products Biochemical Assays P.12 "QuickScout Screening Assist™ Kits" Kinase Protein Assay Kits P.13 "QuickScout Custom Profiling & Panel Profiling Series" Targets P.14 "QuickScout Custom Profiling Series" Preincubation Targets Cell-Based Assays P.15 NanoBRET™ TE Intracellular Kinase Cell-Based Assay Service Targets P.16 Tyrosine Kinase Ba/F3 Cell-Based Assay Service Targets P.17 Kinase HEK293 Cell-Based Assay Service ～ClariCELL™ ～ Targets P.18 Detection of Protein-Protein Interactions ～ProbeX™～ Stable Cell Lines Crystallization Services P.19 FastLane™ Structures ～Premium～ P.20-21 FastLane™ Structures ～Standard～ Kinase Products For details of products, please see "PRODUCTS AND SERVICES" on page 1～3. Tyrosine Kinases Note: Please contact us for availability or further information. Information may be changed without notice. Expression Protein Kinase Tag Carna Product Name Catalog No. Construct Sequence Accession Number Tag Location System HIS ABL(ABL1) 08-001 Full-length 2-1130 NP_005148.2 N-terminal His Insect (sf21) ABL(ABL1) BTN BTN-ABL(ABL1) 08-401-20N Full-length 2-1130 NP_005148.2 N-terminal DYKDDDDK Insect (sf21) ABL(ABL1) [E255K] HIS ABL(ABL1)[E255K] 08-094 Full-length 2-1130 NP_005148.2 N-terminal His Insect (sf21) HIS ABL(ABL1)[T315I] 08-093 Full-length 2-1130 NP_005148.2 N-terminal His Insect (sf21) ABL(ABL1) [T315I] BTN BTN-ABL(ABL1)[T315I] 08-493-20N Full-length 2-1130 NP_005148.2 N-terminal DYKDDDDK Insect (sf21) ACK(TNK2) GST ACK(TNK2) 08-196 Catalytic domain
    [Show full text]
  • Transformation of Human Mesenchymal Stem Cells Increases Their Dependency on Oxidative Phosphorylation for Energy Production
    Transformation of human mesenchymal stem cells increases their dependency on oxidative phosphorylation for energy production Juan M. Funes†‡, Marisol Quintero‡, Stephen Henderson†‡, Dolores Martinez†‡§, Uzma Qureshi¶, Claire Westwood†‡, Mark O. Clements†‡, Dimitra Bourboulia†‡, R. Barbara Pedley¶, Salvador Moncada‡ʈ, and Chris Boshoff†‡ʈ †Cancer Research U.K. Viral Oncology Group and ‡Wolfson Institute for Biomedical Research, Cruciform Building, Gower Street, University College London, London WC 1E 6BT, United Kingdom; and ¶Department of Oncology, Royal Free and University College Medical School, Rowland Hill Street, London NW3 2PF, United Kingdom Contributed by Salvador Moncada, January 25, 2007 (sent for review December 8, 2006) An increased dependency on glycolysis for ATP production is Results considered to be a hallmark of tumor cells. Whether this increase Transformation of Human MSC. Tissue-specific MSC, or mesenchy- in glycolytic activity is due mainly to inherent metabolic alterations mal precursor cells, are thought to be the cell of origin for various or to the hypoxic microenvironment remains controversial. Here types of sarcoma (18). We therefore used human primary MSC we have transformed human adult mesenchymal stem cells (MSC) isolated from bone marrow of a healthy donor. These cells using genetic alterations as described for differentiated cells. Our showed the expected cell surface marker profile (Fig. 1A), and data suggest that MSC require disruption of the same pathways as their multipotency was confirmed by differentiation
    [Show full text]