DOCSLIB.ORG

Proteasome Immunosubunits Protect Against the Development of CD8 T Cell-Mediated Autoimmune Diseases

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Proteasome Immunosubunits Protect Against the Development of CD8 T Cell-Mediated Autoimmune Diseases Proteasome Immunosubunits Protect against the Development of CD8 T Cell-Mediated Autoimmune Diseases This information is current as Dietmar M. W. Zaiss, Cornelis P. J. Bekker, Andrea Gröne, of September 29, 2021. Benedicte A. Lie and Alice J. A. M. Sijts J Immunol published online 29 July 2011 http://www.jimmunol.org/content/early/2011/07/29/jimmun ol.1101003 Downloaded from Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision http://www.jimmunol.org/ • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: by guest on September 29, 2021 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 © 2011 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published July 29, 2011, doi:10.4049/jimmunol.1101003 The Journal of Immunology Proteasome Immunosubunits Protect against the Development of CD8 T Cell-Mediated Autoimmune Diseases Dietmar M. W. Zaiss,* Cornelis P. J. Bekker,* Andrea Gro¨ne,† Benedicte A. Lie,‡ and Alice J. A. M. Sijts* Exposure of cells to inflammatory cytokines induces the expression of three proteasome immunosubunits, two of which are encoded in the MHC class II region. The induced subunits replace their constitutive homologs in newly formed “so-called” immunoproteasomes. Immunosubunit incorporation enhances the proteasome’s proteolytic activity and modifies the proteasome’s cleavage-site preferences, which improves the generation of many MHC class I-presented peptides and shapes the fine specificity of pathogen-specific CD8 T cell responses. In this article, we report on a second effect of immunoproteasome formation on CD8 T cell responses. We show that mice deficient for the immunosubunits b5i/low molecular mass polypeptide (LMP7) and b2i/ multicatalytic endopeptidase complex-like–1 develop early-stage multiorgan autoimmunity following irradiation and bone mar- Downloaded from row transplantation. Disease symptoms are caused by CD8 T cells and are transferable into immunosubunit-deficient, RAG1- deficient mice. Moreover, using the human Type 1 Diabetes Genetics Consortium MHC dataset, we identified two single nucleotide polymorphisms within the b5i/LMP7-encoding gene sequences, which were in strong linkage disequilibrium, as independent genetic risk factors for type 1 diabetes development in humans. Strikingly, these single nucleotide polymorphisms significantly enhanced the risk conferred by HLA haplotypes that were previously shown to predispose for type 1 diabetes. These data suggested that inflammation-induced immunosubunit expression in peripheral tissues constitutes a mechanism that prevents http://www.jimmunol.org/ the development of CD8 T cell-mediated autoimmune diseases. The Journal of Immunology, 2011, 187: 000–000. D8 T cells play an important role in immune protection b2i/ multicatalytic endopeptidase complex-like–1 (MECL-1), and against intracellular pathogens and tumor growth; how- b5i/LMP7 (2, 3), resulting in the formation of immunoprotea- C ever, they also can be mediators of autoimmune disease. somes. CD8 T cells recognize short peptide epitopes that are presented on The three immunosubunits have coevolved with the adaptive the cell surface by MHC class I (MHC-I) molecules. These peptides immune system, and two of these, b1i/LMP2 and b5i/LMP7, are usually are generated upon degradation of intracellular proteins by encoded in the MHC class II (MHC-II) region, suggesting an by guest on September 29, 2021 an abundant protease complex, the proteasome (1). The catalytic important role in immune responses. Indeed, the cleavage site activity of proteasomes is exerted by three subunits, b1, b2, and preferences of immunoproteasomes differ from those of consti- b5, which are constitutively expressed in all cells. In hemato- tutive proteasomes; consequently, the repertoire of peptides pro- poietic cells and cells exposed to inflammatory cytokines, such as duced by immunoproteasomes differs from that produced by TNF-a and IFN-g, these subunits are exchanged for three cytokine- constitutive proteasome complexes (4, 5). These qualitative dif- inducible homologs, b1i/low molecular mass polypeptide (LMP)2, ferences in peptide generation are reflected in the immunodomi- nance hierarchy of pathogen-specific CD8 T cell responses, thus several epitopes that are dominant targets of the pathogen- *Division of Immunology, Faculty of Veterinary Medicine, University of Utrecht, specific CD8 T cell response in infected wild type (wt) mice are † 3584CL Utrecht, The Netherlands; Division of Pathology, Faculty of Veterinary not targeted by CD8 T cells in immunosubunit-deficient mice due Medicine, University of Utrecht, 3584CL Utrecht, The Netherlands; and ‡Institute of Immunology, Oslo University Hospital Rikshospitalet, N-0424 Oslo, Norway to inefficient generation (6, 7). However, CD8 T cell responses Received for publication April 6, 2011. Accepted for publication June 21, 2011. to immunoproteasome-generated epitopes are not essential for clearance of most pathogens (5, 8). Recent studies indicated that, This work was supported by National Institutes of Health Grant AI064576 (to A.J.A.M.S.) and the MEERVOUD program of The Netherlands Organisation for under conditions of IFN-induced oxidative stress, immunopro- Scientific Research. This work used resources provided by the Type 1 Diabetes teasomes could play a more general role in the maintenance of Genetics Consortium, a collaborative clinical study sponsored by the National In- stitute of Diabetes and Digestive Kidney Diseases, National Allergy and Infectious protein homeostasis (9). Whereas IFN-exposed immunoproteasome- Diseases, National Human Genome Research Institute, National Institute of Child deficient cells were found to accumulate oxidant-damaged, poly- Health and Human Development, and the Juvenile Diabetes Research Foundation ubiquinated unfolded nascent proteins in aggresome-like induced International and supported by National Institutes of Health Grant U01 DK062418. structures, formation of such structures was only transient in cells Address correspondence and reprint requests to Dr. A. Sijts, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, University of Utrecht, with immunoproteasomes, as a result of the enhanced proteolysis. Yalelaan 1, 3584CL Utrecht, The Netherlands. E-mail address: [email protected] Recently, it has become more appreciated that, in addition to Abbreviations used in this article: BM, bone marrow; CI, confidence interval; IDDM, CD4 T cells, the CD8 T cell subset plays an important role in tissue insulin-dependent diabetes mellitus; IGRP, islet-specific glucose-6-phosphatase cat- destruction during the progression of autoimmune disease (10–13). alytic subunit related protein; LD, linkage disequilibrium; LMP, low molecular mass polypeptide; MECL-1, multicatalytic endopeptidase complex-like–1; MHC-I, MHC In this article, we show that mice, gene-deficient for b2i/MECL-1 class I; MHC-II, MHC class II; o/n, overnight; OR, odds ratio; SNP, single nucleotide and b5i/LMP7, develop CD8 T cell-mediated, early-stage, mul- polymorphism; T1D, type 1 diabetes; T1DGC, Type-1 Diabetes Genetics Consor- titissue autoimmune syndrome after irradiation and bone marrow tium; UTR, untranslated region; wt, wild type. (BM) reconstitution. These data indicate that one important Copyright Ó 2011 by The American Association of Immunologists, Inc. 0022-1767/11/$16.00 function of the immunosubunits, as well as their inducible www.jimmunol.org/cgi/doi/10.4049/jimmunol.1101003 2 IMMUNOPROTEASOMES PROTECT AGAINST AUTOIMMUNE DISEASES expression in inflamed tissues, is the prevention of CD8 T cell- coding of DRB1, DQA1, and DQB1 haplotypes, was described previously mediated autoimmune reactions. (15). IFN-g–ELISPOT Materials and Methods Ninety-six–well ELISPOT plates (Milipore, Billerica, MA) were coated with 2 mg/ml AN18 (anti-mouse IFN-g) in 100 ml PBS for $2 h at room Mice temperature. Wells were then washed and blocked twice with RPMI 1640 3 6 C57BL/6 (B6), B6 RAG1-, and b5i/LMP7 and b2i/MECL-1 gene-deficient medium. Splenocyte dilutions, starting with 1 10 cells/well, were in- mice (14), backcrossed .10 times onto the B6 background, were maintained cubated for $4 h in the presence or absence of 2 mg/ml synthetic peptide by in-house breeding under specific pathogen-free conditions. RAG12/2 epitopes in 100 ml IMDM medium. Thereafter, the plates were washed mice were crossed with b5i/LMP7 and b2i/MECL-1 gene-deficient mice to with PBS plus 0.01% Tween 20, and IFN-g was detected by incuba- generate b5i/LMP7 and RAG1 gene-deficient mice. PSMB8 deficiency was tion with 2 mg/ml biotinylated XMG1.2, followed by 1 mg/ml alkaline detected by PCR, using the oligonucleotides 59-GGACCAGGACTTTAC- phosphatase-conjugated streptavidin (Jackson Immunobiology, Bar Har- TACGTAGATG-39 on PSMB8 or 59-CCGACGGCGAGGATCTCGTCG- bor, ME) in PBS plus 0.01% Tween 20 supplemented with 2% BSA. The TGA-39 on the neomycin cassette as forward primer and the PSMB8-
Load more

Recommended publications
  • Genetic Variations in the PSMA6 and PSMC6 Proteasome Genes Are Associated with Multiple Sclerosis and Response to Interferon‑Β Therapy in Latvians
    EXPERIMENTAL AND THERAPEUTIC MEDICINE 21: 478, 2021 Genetic variations in the PSMA6 and PSMC6 proteasome genes are associated with multiple sclerosis and response to interferon‑β therapy in Latvians NATALIA PARAMONOVA1, JOLANTA KALNINA1, KRISTINE DOKANE1, KRISTINE DISLERE1, ILVA TRAPINA1, TATJANA SJAKSTE1 and NIKOLAJS SJAKSTE1,2 1Genomics and Bioinformatics, Institute of Biology of The University of Latvia; 2Department of Medical Biochemistry of The University of Latvia, LV‑1004 Riga, Latvia Received July 8, 2020; Accepted December 8, 2020 DOI: 10.3892/etm.2021.9909 Abstract. Several polymorphisms in genes related to the Introduction ubiquitin‑proteasome system exhibit an association with pathogenesis and prognosis of various human autoimmune Multiple sclerosis (MS) is a lifelong demyelinating disease of diseases. Our previous study reported the association the central nervous system. The clinical onset of MS tends to between multiple sclerosis (MS) and the PSMA3‑rs2348071 be between the second and fourth decade of life. Similarly to polymorphism in the Latvian population. The current study other autoimmune diseases, women are affected 3‑4 times more aimed to evaluate the PSMA6 and PSMC6 genetic variations, frequently than men (1). About 10% of MS patients experience their interaction between each other and with the rs2348071, a primary progressive MS form characterized by the progres‑ on the susceptibility to MS risk and response to therapy in sion of neurological disability from the onset. In about 90% the Latvian population. PSMA6‑rs2277460, ‑rs1048990 and of MS patients, the disease undergoes the relapse‑remitting PSMC6‑rs2295826, ‑rs2295827 were genotyped in the MS MS course (RRMS); in most of these patients, the condition case/control study and analysed in terms of genotype‑protein acquires secondary progressive course (SPMS) (2).
    [Show full text]
  • An Integrative Analysis of Gene Expression and Molecular
    Muraro and Simmons BMC Bioinformatics (2016) 17:42 DOI 10.1186/s12859-016-0886-z RESEARCH ARTICLE Open Access An integrative analysis of gene expression and molecular interaction data to identify dys-regulated sub-networks in inflammatory bowel disease Daniele Muraro* and Alison Simmons Abstract Background: Inflammatory bowel disease (IBD) consists of two main disease-subtypes, Crohn’s disease (CD) and ulcerative colitis (UC); these subtypes share overlapping genetic and clinical features. Genome-wide microarray data enable unbiased documentation of alterations in gene expression that may be disease-specific. As genetic diseases are believed to be caused by genetic alterations affecting the function of signalling pathways, module-centric optimisation algorithms, whose aim is to identify sub-networks that are dys-regulated in disease, are emerging as promising approaches. Results: In order to account for the topological structure of molecular interaction networks, we developed an optimisation algorithm that integrates databases of known molecular interactions with gene expression data; such integration enables identification of differentially regulated network modules. We verified the performance of our algorithm by testing it on simulated networks; we then applied the same method to study experimental data derived from microarray analysis of CD and UC biopsies and human interactome databases. This analysis allowed the extraction of dys-regulated subnetworks under different experimental conditions (inflamed and uninflamed tissues in CD and UC). Optimisation was performed to highlight differentially expressed network modules that may be common or specific to the disease subtype. Conclusions: We show that the selected subnetworks include genes and pathways of known relevance for IBD; in particular, the solutions found highlight cross-talk among enriched pathways, mainly the JAK/STAT signalling pathway and the EGF receptor signalling pathway.
    [Show full text]
  • Protein Expression Analysis of an in Vitro Murine Model of Prostate Cancer Progression: Towards Identification of High-Potential Therapeutic Targets
    Journal of Personalized Medicine Article Protein Expression Analysis of an In Vitro Murine Model of Prostate Cancer Progression: Towards Identification of High-Potential Therapeutic Targets Hisham F. Bahmad 1,2,3 , Wenjing Peng 4, Rui Zhu 4, Farah Ballout 1, Alissar Monzer 1, 1,5 6, , 1, , 4, , Mohamad K. Elajami , Firas Kobeissy * y , Wassim Abou-Kheir * y and Yehia Mechref * y 1 Department of Anatomy, Cell Biology and Physiological Sciences, Faculty of Medicine, American University of Beirut, Beirut 1107-2020, Lebanon; [email protected] (H.F.B.); [email protected] (F.B.); [email protected] (A.M.); [email protected] (M.K.E.) 2 Arkadi M. Rywlin M.D. Department of Pathology and Laboratory Medicine, Mount Sinai Medical Center, Miami Beach, FL 33140, USA 3 Herbert Wertheim College of Medicine, Florida International University, Miami, FL 33199, USA 4 Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA; [email protected] (W.P.); [email protected] (R.Z.) 5 Department of Internal Medicine, Mount Sinai Medical Center, Miami Beach, FL 33140, USA 6 Department of Biochemistry and Molecular Genetics, Faculty of Medicine, American University of Beirut, Beirut 1107-2020, Lebanon * Correspondence: [email protected] (F.K.); [email protected] (W.A.-K.); [email protected] (Y.M.); Tel.: +961-1-350000 (ext. 4805) (F.K.); +961-1-350000 (ext. 4778) (W.A.K.); +1-806-834-8246 (Y.M.); Fax: +1-806-742-1289 (Y.M.); 961-1-744464 (W.A.K.) These authors have contributed equally to this work as joint senior authors.
    [Show full text]
  • Molecular Pathology and Novel Clinical Therapy for Uterine
    ANTICANCER RESEARCH 36 : 4997-5008 (2016) doi:10.21873/anticanres.11068 Review Molecular Pathology and Novel Clinical Therapy for Uterine Leiomyosarcoma TAKUMA HAYASHI 1,2 , MIKI KAWANO 2,3 , TOMOYUKI ICHIMURA 4, KOICHI IDA 1, HIROFUMI ANDO 1, DORIT ZHARHARY 5, YAE KANAI 6, HIROYUKI ABURATANI 7, SUSUMU TONEGAWA 8, TANRI SHIOZAWA 1, NOBUO YAEGASHI 9 and IKUO KONISHI 10 1Department of Obstetrics and Gynecology, Shinshu University School of Medicine, Nagano, Japan; 2Department of Medical Technology, International University of Health and Welfare, Chiba, Japan; 3Department of Health Science, Kyushu University Graduate School of Medicine, Fukuoka, Japan; 4Department of Obstetrics and Gynecology, Osaka City University Graduate School of Medicine, Osaka, Japan; 5SIGMA-Aldrich Israel, Rehovot, Israel; 6Pathology Division, Keio University School of Medicine, Tokyo, Japan; 7The Cancer System Laboratory, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan; 8Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, U.S.A.; 9Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, Miyagi, Japan; 10 National Hospital Organization Kyoto Medical Centre, Kyoto, Japan Abstract. Patients with uterine leiomyosarcoma (LMS) disease prevalence of ~37% by 12 months of age. Furthermore, typically present with vaginal bleeding, pain, and a pelvic a recent report showed the loss of ability to induce PSMB9/ β1 i mass, with atypical presentations of hypercalcemia and expression,
    [Show full text]
  • Micrornas in the Etiology of Colorectal Cancer: Pathways and Clinical Implications Ashlee M
    Washington University School of Medicine Digital Commons@Becker Open Access Publications 2017 MicroRNAs in the etiology of colorectal cancer: Pathways and clinical implications Ashlee M. Strubberg Washington University School of Medicine in St. Louis Blair B. Madison Washington University School of Medicine in St. Louis Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Recommended Citation Strubberg, Ashlee M. and Madison, Blair B., ,"MicroRNAs in the etiology of colorectal cancer: Pathways and clinical implications." Disease Models & Mechanisms.10,3. 197-214. (2017). https://digitalcommons.wustl.edu/open_access_pubs/5644 This Open Access Publication is brought to you for free and open access by Digital Commons@Becker. It has been accepted for inclusion in Open Access Publications by an authorized administrator of Digital Commons@Becker. For more information, please contact [email protected]. © 2017. Published by The Company of Biologists Ltd | Disease Models & Mechanisms (2017) 10, 197-214 doi:10.1242/dmm.027441 REVIEW MicroRNAs in the etiology of colorectal cancer: pathways and clinical implications Ashlee M. Strubberg and Blair B. Madison* ABSTRACT rates from colorectal cancer have declined over the past 20 years, MicroRNAs (miRNAs) are small single-stranded RNAs that repress largely thanks to early detection; however, CRC incidence still mRNA translation and trigger mRNA degradation. Of the ∼1900 remains high (Siegel et al., 2015) and new treatments have been miRNA-encoding genes present in the human genome, ∼250 lagging. Treatment usually entails surgical removal of tumors, miRNAs are reported to have changes in abundance or altered which may be followed with chemotherapy and/or targeted functions in colorectal cancer.
    [Show full text]
  • Datasheet: VPA00155 Product Details
    Datasheet: VPA00155 Description: GOAT ANTI PSMB8 Specificity: PSMB8 Format: Purified Product Type: PrecisionAb™ Polyclonal Isotype: Polyclonal IgG Quantity: 100 µl Product Details Applications This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit www.bio-rad-antibodies.com/protocols. Yes No Not Determined Suggested Dilution Western Blotting 1/1000 PrecisionAb antibodies have been extensively validated for the western blot application. The antibody has been validated at the suggested dilution. Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Further optimization may be required dependant on sample type. Target Species Human Species Cross Reacts with: Rat Reactivity N.B. Antibody reactivity and working conditions may vary between species. Product Form Purified IgG - liquid Preparation Goat polyclonal antibody purified by affinity chromatography Buffer Solution TRIS buffered saline Preservative 0.02% Sodium Azide (NaN ) 0.5 % BSA Stabilisers 3 Immunogen Peptide with the sequence C-DVSDLLHQYREANQ, from the C terminus of the protein sequence External Database Links UniProt: P28062 Related reagents Entrez Gene: 5696 PSMB8 Related reagents Synonyms LMP7, PSMB5i, RING10, Y2 Page 1 of 2 Specificity Goat anti Human PSMB8 antibody recognizes proteasome subunit beta type-8, also known as Low molecular mass protein 7, macropain subunit C13, multicatalytic endopeptidase complex subunit C13, proteasome component C13, and proteasome subunit beta-5i. The proteasome is a multicatalytic proteinase complex with a highly ordered ring-shaped 20S core structure.
    [Show full text]
  • Tyrosine Kinase Chromosomal Translocations Mediate Distinct and Overlapping Gene Regulation Events
    Kim et al. BMC Cancer 2011, 11:528 http://www.biomedcentral.com/1471-2407/11/528 RESEARCHARTICLE Open Access Tyrosine kinase chromosomal translocations mediate distinct and overlapping gene regulation events Hani Kim1,3, Lisa C Gillis1,2, Jordan D Jarvis1,2, Stuart Yang3, Kai Huang1, Sandy Der3 and Dwayne L Barber1,2,3* Abstract Background: Leukemia is a heterogeneous disease commonly associated with recurrent chromosomal translocations that involve tyrosine kinases including BCR-ABL, TEL-PDGFRB and TEL-JAK2. Most studies on the activated tyrosine kinases have focused on proximal signaling events, but little is known about gene transcription regulated by these fusions. Methods: Oligonucleotide microarray was performed to compare mRNA changes attributable to BCR-ABL, TEL- PDGFRB and TEL-JAK2 after 1 week of activation of each fusion in Ba/F3 cell lines. Imatinib was used to control the activation of BCR-ABL and TEL-PDGFRB, and TEL-JAK2-mediated gene expression was examined 1 week after Ba/F3- TEL-JAK2 cells were switched to factor-independent conditions. Results: Microarray analysis revealed between 800 to 2000 genes induced or suppressed by two-fold or greater by each tyrosine kinase, with a subset of these genes commonly induced or suppressed among the three fusions. Validation by Quantitative PCR confirmed that eight genes (Dok2, Mrvi1, Isg20, Id1, gp49b, Cxcl10, Scinderin, and collagen Va1(Col5a1)) displayed an overlapping regulation among the three tested fusion proteins. Stat1 and Gbp1 were induced uniquely by TEL-PDGFRB. Conclusions: Our results suggest that BCR-ABL, TEL-PDGFRB and TEL-JAK2 regulate distinct and overlapping gene transcription profiles.
    [Show full text]
  • Intratumoral Injection of SYNB1891, a Synthetic Biotic Medicine Designed
    Intratumoral injection of SYNB1891 A Synthetic Biotic medicine designed to activate the innate immune system. Therapy demonstrates target engagement in humans including intratumoral STING activation. Janku F, MD Anderson Cancer Center; Luke JJ, UPMC Hillman Cancer Center; Brennan AM, Synlogic; Riese RJ, Synlogic; Varterasian M, Pharmaceutical Consultant; Kuhn K, Synlogic; Sokolovska A, Synlogic; Strauss J, Mary Crowley Cancer Research Presented by Filip Janku, MD, PhD Study supported by Synlogic, Inc American Association for Cancer Research (AACR) April 2021 Introduction and Methods SYNB1891 Strain Phase 1 First-in-Human Clinical Trial • Live, modified strain of the probiotic E. coli • Enrolling patients with refractory advanced solid Nissle engineered to produce cyclic tumors or lymphoma dinucleotides (CDN) under hypoxia leading to stimulator of interferon genes (STING)- • Intratumoral (IT) injection of SYNB1891 on Days activation 1, 8 and 15 of the first 21-day cycle and then on Day 1 of each subsequent cycle. • Preferentially taken up by phagocytic antigen- presenting cells in tumors, activating • Dose escalation planned across 7 cohorts (1x106 complementary innate immune pathways – 1x109 live cells) with Arm 1 consisting of (direct CDN STING activation; cGAS-mediated SYNB1891 as monotherapy, and Arm 2 in STING activation and TLR4/MyD88 activation by combination with atezolizumab the bacterial chassis) SYNB1891 was safe and well-tolerated in heterogenous population Nov 2020: Interim Analysis IA Updated through 15 Mar 2021 15 Mar 2021:
    [Show full text]
  • Biological Models of Colorectal Cancer Metastasis and Tumor Suppression
    BIOLOGICAL MODELS OF COLORECTAL CANCER METASTASIS AND TUMOR SUPPRESSION PROVIDE MECHANISTIC INSIGHTS TO GUIDE PERSONALIZED CARE OF THE COLORECTAL CANCER PATIENT By Jesse Joshua Smith Dissertation Submitted to the Faculty of the Graduate School of Vanderbilt University In partial fulfillment of the requirements For the degree of DOCTOR OF PHILOSOPHY In Cell and Developmental Biology May, 2010 Nashville, Tennessee Approved: Professor R. Daniel Beauchamp Professor Robert J. Coffey Professor Mark deCaestecker Professor Ethan Lee Professor Steven K. Hanks Copyright 2010 by Jesse Joshua Smith All Rights Reserved To my grandparents, Gladys and A.L. Lyth and Juanda Ruth and J.E. Smith, fully supportive and never in doubt. To my amazing and enduring parents, Rebecca Lyth and Jesse E. Smith, Jr., always there for me. .my sure foundation. To Jeannine, Bill and Reagan for encouragement, patience, love, trust and a solid backing. To Granny George and Shawn for loving support and care. And To my beautiful wife, Kelly, My heart, soul and great love, Infinitely supportive, patient and graceful. ii ACKNOWLEDGEMENTS This work would not have been possible without the financial support of the Vanderbilt Medical Scientist Training Program through the Clinical and Translational Science Award (Clinical Investigator Track), the Society of University Surgeons-Ethicon Scholarship Fund and the Surgical Oncology T32 grant and the Vanderbilt Medical Center Section of Surgical Sciences and the Department of Surgical Oncology. I am especially indebted to Drs. R. Daniel Beauchamp, Chairman of the Section of Surgical Sciences, Dr. James R. Goldenring, Vice Chairman of Research of the Department of Surgery, Dr. Naji N.
    [Show full text]
  • Additive Loss-Of-Function Proteasome Subunit Mutations in CANDLE/PRAAS Patients Promote Type I IFN Production
    Amendment history: Erratum (February 2016) Additive loss-of-function proteasome subunit mutations in CANDLE/PRAAS patients promote type I IFN production Anja Brehm, … , Ivona Aksentijevich, Raphaela Goldbach-Mansky J Clin Invest. 2015;125(11):4196-4211. https://doi.org/10.1172/JCI81260. Research Article Immunology Autosomal recessive mutations in proteasome subunit β 8 (PSMB8), which encodes the inducible proteasome subunit β5i, cause the immune-dysregulatory disease chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature (CANDLE), which is classified as a proteasome-associated autoinflammatory syndrome (PRAAS). Here, we identified 8 mutations in 4 proteasome genes, PSMA3 (encodes α7), PSMB4 (encodes β7), PSMB9 (encodes β1i), and proteasome maturation protein (POMP), that have not been previously associated with disease and 1 mutation inP SMB8 that has not been previously reported. One patient was compound heterozygous for PSMB4 mutations, 6 patients from 4 families were heterozygous for a missense mutation in 1 inducible proteasome subunit and a mutation in a constitutive proteasome subunit, and 1 patient was heterozygous for a POMP mutation, thus establishing a digenic and autosomal dominant inheritance pattern of PRAAS. Function evaluation revealed that these mutations variably affect transcription, protein expression, protein folding, proteasome assembly, and, ultimately, proteasome activity. Moreover, defects in proteasome formation and function were recapitulated by siRNA-mediated knockdown of the respective
    [Show full text]
  • PSMB9/LMP2 Rabbit Pab
    Leader in Biomolecular Solutions for Life Science PSMB9/LMP2 Rabbit pAb Catalog No.: A1771 1 Publications Basic Information Background Catalog No. The proteasome is a multicatalytic proteinase complex with a highly ordered ring- A1771 shaped 20S core structure. The core structure is composed of 4 rings of 28 non-identical subunits; 2 rings are composed of 7 alpha subunits and 2 rings are composed of 7 beta Observed MW subunits. Proteasomes are distributed throughout eukaryotic cells at a high 22kDa concentration and cleave peptides in an ATP/ubiquitin-dependent process in a non- lysosomal pathway. An essential function of a modified proteasome, the Calculated MW immunoproteasome, is the processing of class I MHC peptides. This gene encodes a 22kDa/23kDa member of the proteasome B-type family, also known as the T1B family, that is a 20S core beta subunit. This gene is located in the class II region of the MHC (major Category histocompatibility complex). Expression of this gene is induced by gamma interferon and this gene product replaces catalytic subunit 1 (proteasome beta 6 subunit) in the Primary antibody immunoproteasome. Proteolytic processing is required to generate a mature subunit. Applications WB, IF Cross-Reactivity Human, Mouse Recommended Dilutions Immunogen Information WB 1:500 - 1:2000 Gene ID Swiss Prot 5698 P28065 IF 1:10 - 1:100 Immunogen Recombinant fusion protein containing a sequence corresponding to amino acids 1-219 of human PSMB9/LMP2/LMP2 (NP_002791.1). Synonyms PSMB9;LMP2;PSMB6i;RING12;beta1i Contact Product Information www.abclonal.com Source Isotype Purification Rabbit IgG Affinity purification Storage Store at -20℃.
    [Show full text]
  • PSMB8 Gene Proteasome Subunit Beta 8
    PSMB8 gene proteasome subunit beta 8 Normal Function The PSMB8 gene provides instructions for making one part (subunit) of cell structures called immunoproteasomes. Immunoproteasomes are specialized versions of proteasomes, which are large complexes that recognize and break down (degrade) unneeded, excess, or abnormal proteins within cells. This activity is necessary for many essential cell functions. While proteasomes are found in many types of cells, immunoproteasomes are located primarily in immune system cells. These structures play an important role in regulating the immune system's response to foreign invaders, such as viruses and bacteria. One of the primary functions of immunoproteasomes is to help the immune system distinguish the body's own proteins from proteins made by foreign invaders, so the immune system can respond appropriately to infection. Immunoproteasomes may also have other functions in immune system cells and possibly in other types of cells. They appear to be involved in some of the same fundamental cell activities as regular proteasomes, such as regulating the amount of various proteins in cells (protein homeostasis), cell growth and division, the process by which cells mature to carry out specific functions (differentiation), chemical signaling within cells, and the activity of genes. Studies suggest that, through unknown mechanisms, the subunit produced from the PSMB8 gene in particular may be involved in the maturation of fat cells (adipocytes). Health Conditions Related to Genetic Changes Nakajo-Nishimura syndrome At least one mutation in the PSMB8 gene has been found to cause Nakajo-Nishimura syndrome, a condition that has been described only in the Japanese population. The identified mutation changes a single protein building block (amino acid) in the protein produced from the PSMB8 gene, replacing the amino acid glycine with the amino acid valine at protein position 201 (written as Gly201Val or G201V).
    [Show full text]