DOCSLIB.ORG

The Oncogenic Role of the Protein Tyrosine Phosphatase 4A3 (Ptp4a3 Or Prl-3) in T-Cell Acute Lymphoblastic Leukemia

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Oncogenic Role of the Protein Tyrosine Phosphatase 4A3 (Ptp4a3 Or Prl-3) in T-Cell Acute Lymphoblastic Leukemia University of Kentucky UKnowledge UKnowledge Theses and Dissertations--Molecular and Cellular Biochemistry Molecular and Cellular Biochemistry 2020 THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA Min Wei University of Kentucky, [email protected] Author ORCID Identifier: https://orcid.org/0000-0001-5112-7288 Digital Object Identifier: https://doi.org/10.13023/etd.2020.278 Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Wei, Min, "THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA" (2020). Theses and Dissertations--Molecular and Cellular Biochemistry. 46. https://uknowledge.uky.edu/biochem_etds/46 This Doctoral Dissertation is brought to you for free and open access by the Molecular and Cellular Biochemistry at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Molecular and Cellular Biochemistry by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Min Wei, Student Dr. Jessica Blackburn, Major Professor Dr. Trevor Creamer, Director of Graduate Studies University of Kentucky UKnowledge Theses and Dissertations--Molecular and Cellular Biochemistry Molecular and Cellular Biochemistry 2020 THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA Min Wei Right click to open a feedback form in a new tab to let us know how this document benefits ou.y STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Min Wei, Student Dr. Jessica Blackburn, Major Professor Dr. Trevor Creamer, Director of Graduate Studies THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA ________________________________________ DISSERTATION ________________________________________ A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the College of Medicine at the University of Kentucky By Min Wei Lexington, Kentucky Co-Director: Dr. Jessica Blackburn, Assistant Professor of Biochemistry and Dr.Tianyan Gao, Professor of Biochemistry Lexington, Kentucky 2020 Copyright © Min Wei 2020 [https://orcid.org/0000-0001-5112-7288] ABSTRACT OF DISSERTATION THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA T-cell Acute Lymphoblastic Leukemia (T-ALL) is an aggressive blood cancer. There are no immunotherapies and few molecularly targeted therapeutics available for the treatment of this malignancy. The identification and characterization of genes and pathways that drive T-ALL progression is critical for the development of new therapies for T-ALL. The protein tyrosine phosphatase PTP4A3, also known as PRL-3, has been extensively reported to play a causative role in numerous cancers, including several types of blood malignancies. However, its role in T-ALL is not well defined. Here, we determined that the PRL-3 plays a critical role in T-ALL initiation and progression by promoting leukemia cell migration. PRL-3 is highly expressed in patient T- ALL samples at both mRNA and protein levels compared to normal lymphocytes. Knock- down of PRL-3 expression using short-hairpin RNA (shRNA) in human T-ALL cell lines significantly impeded T-ALL cell migration capacity in vitro and reduced their ability to engraft and proliferate in xenograft mouse models. Additionally, PRL-3 over-expression in a Myc-induced zebrafish T-ALL model significantly accelerated disease onset and shortened the time needed for cells to enter blood circulation. Reverse phase protein array (RPPA) and gene set enrichment analysis (GSEA) revealed that the SRC signaling pathway is affected by PRL-3. Immunoblot analyses validated that manipulation of PRL-3 expression in T-ALL cells affected the SRC signaling pathway, which is directly involved in cell migration, although SRC was not a direct substrate of PRL-3. More importantly, T- ALL cell growth and migration was inhibited by small molecule inhibition of PRL-3, suggesting that PRL-3 has potential as a therapeutic target in T-ALL. Finally, we identified a pool of PRL-3 interacting proteins in T-ALL using BioID- based proximal labeling approach. Co-immunoprecipitation assays were used to validate a number of direct interacting partners of PRL-3, including LCK and CD3. Taken together, our study identifies PRL-3 as an oncogenic driver in T-ALL both in vitro and in vivo and provides a strong rationale for targeted therapies that interfere with PRL-3 function. We also identified PRL-3 interactomes in T-ALL, which lays foundation for future PRL-3 substrates study. KEYWORDS: PRL3, T-ALL, Zebrafish, Migration, BioID2, Proximal-labeling Min Wei (Name of Student) [06/10/2020] Date THE ONCOGENIC ROLE OF THE PROTEIN TYROSINE PHOSPHATASE 4A3 (PTP4A3 OR PRL-3) IN T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA By Min Wei Jessica Blackburn Co-Director of Dissertation Tianyan Gao Co-Director of Dissertation Trevor Creamer Director of Graduate Studies [06/10/2020] Date ACKNOWLEDGMENTS There are so many people I need to thank during my research and life in the past 5 years. First and foremost, I would like to express heartiest gratitude and sincere thanks to my mentor, Dr. Jessica Blackburn. Not only for her persistent guidance and help throughout every step of my research, but also for that she has continually conveyed a spirit of caring, kindness, and trust for people working with her and demonstrated the power of gentleness, which will greatly impact my life. I have seen how much effort she put in to help trainees to grow and achieve success, which is a great asset to the science community. I very much appreciate the current and past members of Blackburn lab who created the collegial atmosphere of our lab and have made my time here rememberable. Particularly I would like to say thank you Meghan Haney and Caroline Smith. They are not only great people to work with but also people who can share with you up and downs in life. I appreciate Dylan Rivas’ help for sharing his unique and thoughtful perspective about experimental design. I want to thank Yelena Chernyavskaya, Shaw Powell, Mary Wimsett, Sergei Revskoy and Henry Moore for their friendship and general support. I would also like to extend my deepest gratitude to my dissertation committee members, Dr. Tianyan Gao, Dr. Emilia Galperin, and Dr. John A. D'Orazio for providing me valuable guidance and regular feedback. I really appreciate the time they spent and effort they devoted to shape my dissertation project throughout this study. Also, I would like to thank the outside examiner, Dr. Ernest Snow for his valuable time, knowledge, and feedback that is really helpful to my dissertation improvement and defense.
Load more

Recommended publications
  • Clinical and Biological Features of PTPN2-Deleted Adult and Pediatric T-Cell Acute Lymphoblastic Leukemia
    REGULAR ARTICLE Clinical and biological features of PTPN2-deleted adult and pediatric T-cell acute lymphoblastic leukemia Marion Alcantara,1 Mathieu Simonin,1-3 Ludovic Lhermitte,1 Aurore Touzart,1 Marie Emilie Dourthe,1,4 Mehdi Latiri,1 Nathalie Grardel,5 Jean Michel Cayuela,6 Yves Chalandon,7 Carlos Graux,8 Herve´ Dombret,9 Norbert Ifrah,10 Arnaud Petit,2,3 Elizabeth Macintyre,1 Andre´ Baruchel,4 Nicolas Boissel,9 and Vahid Asnafi1 1Universite´ Paris Descartes Sorbonne Cite,´ Institut Necker-Enfants Malades, Institut National de Recherche Medicale´ U1151, Laboratory of Onco-Hematology, Assistance Publique–Hopitauxˆ de Paris, Hopitalˆ Necker Enfants-Malades, Paris, France; 2Department of Pediatric Hematology and Oncology, Assistance Publique–Hopitauxˆ de Paris, Groupe Hospitalier des Hopitauxˆ Universitaires Est Parisien, Armand Trousseau Hospital, Paris, France; 3Sorbonne Universites,´ Universite´ Pierre et Marie Curie Universite´ Paris 06, Unite´ Mixte de Recherche S 938, Centre de Recherche Saint-Antoine, Groupe de Recherche (GRC) Clinique n°07, GRC Myeloprolif´ erations´ aigues¨ et chroniques, Paris, France; 4Department of Pediatric Hematology and Immunology, Assistance Publique–Hopitauxˆ de Paris, Hopitalˆ Universitaire Robert-Debre,´ Paris, France; 5Laboratory of Hematology, Centre Hospitalier Regional´ Universitaire de Lille, Lille, France; 6Laboratory of Hematology, Saint-Louis Hospital, Assistance Publique–Hopitauxˆ de Paris, Paris, France; 7Swiss Group for Clinical Cancer Research, Bern, Hopitalˆ Universitaire, Geneva, Switzerland; 8Universite´ Catholique de Louvain, Centre Hospitalier Universitaire, Namur, Yvoir, Belgium; 9Adolescent and Young Adult Hematology Unit, Saint-Louis Hospital, Assistance Publique–Hopitauxˆ de Paris, Paris, France; and 10Centre Hospitalier Universitaire d’Angers, Angers, France Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a phosphatase known to be Key Points a tumor suppressor gene in T-cell acute lymphoblastic leukemia (T-ALL).
    [Show full text]
  • Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase As a Molecular Target for Ovarian Cancer
    biomolecules Article Credentialing and Pharmacologically Targeting PTP4A3 Phosphatase as a Molecular Target for Ovarian Cancer John S. Lazo 1,2,* , Elizabeth R. Sharlow 1,2,*, Robert Cornelison 1,2, Duncan J. Hart 1, Danielle C. Llaneza 1, Anna J. Mendelson 1, Ettore J. Rastelli 3 , Nikhil R. Tasker 3, Charles N. Landen, Jr. 4 and Peter Wipf 3 1 Department of Pharmacology, University of Virginia, Charlottesville, VA 22908, USA; [email protected] (R.C.); [email protected] (D.J.H.); [email protected] (D.C.L.); [email protected] (A.J.M.) 2 KeViRx, Inc., Charlottesville, VA 22904, USA 3 Department of Chemistry, University of Pittsburgh, Pittsburgh, PA 15260, USA; [email protected] (E.J.R.); [email protected] (N.R.T.); [email protected] (P.W.) 4 Department of Obstetrics and Gynecology, University of Virginia, Charlottesville, VA 22908, USA; [email protected] * Correspondence: [email protected] or [email protected] (J.S.L.); [email protected] (E.R.S.); Tel.: +1-434-243-1936 (J.S.L.); +1-434-243-1937 (E.R.S.) Abstract: High grade serous ovarian cancer (OvCa) frequently becomes drug resistant and often recurs. Consequently, new drug targets and therapies are needed. Bioinformatics-based studies uncovered a relationship between high Protein Tyrosine Phosphatase of Regenerating Liver-3 (PRL3 also known as PTP4A3) expression and poor patient survival in both early and late stage OvCa. PTP4A3 mRNA levels were 5–20 fold higher in drug resistant or high grade serous OvCa cell lines Citation: Lazo, J.S.; Sharlow, E.R.; compared to nonmalignant cells. JMS-053 is a potent allosteric small molecule PTP4A3 inhibitor Cornelison, R.; Hart, D.J.; Llaneza, and to explore further the role of PTP4A3 in OvCa, we synthesized and interrogated a series of D.C.; Mendelson, A.J.; Rastelli, E.J.; JMS-053-based analogs in OvCa cell line-based phenotypic assays.
    [Show full text]
  • Inflammatory Cytokine Signalling by Protein Tyrosine Phosphatases in Pancreatic Β-Cells
    59 4 W J STANLEY and others PTPN1 and PTPN6 modulate 59: 4 325–337 Research cytokine signalling in β-cells Differential regulation of pro- inflammatory cytokine signalling by protein tyrosine phosphatases in pancreatic β-cells William J Stanley1,2, Prerak M Trivedi1,2, Andrew P Sutherland1, Helen E Thomas1,2 and Esteban N Gurzov1,2,3 Correspondence should be addressed 1 St. Vincent’s Institute of Medical Research, Melbourne, Australia to E N Gurzov 2 Department of Medicine, St. Vincent’s Hospital, The University of Melbourne, Melbourne, Australia Email 3 ULB Center for Diabetes Research, Universite Libre de Bruxelles (ULB), Brussels, Belgium esteban.gurzov@unimelb. edu.au Abstract Type 1 diabetes (T1D) is characterized by the destruction of insulin-producing β-cells Key Words by immune cells in the pancreas. Pro-inflammatory including TNF-α, IFN-γ and IL-1β f pancreatic β-cells are released in the islet during the autoimmune assault and signal in β-cells through f protein tyrosine phosphorylation cascades, resulting in pro-apoptotic gene expression and eventually phosphatases β-cell death. Protein tyrosine phosphatases (PTPs) are a family of enzymes that regulate f PTPN1 phosphorylative signalling and are associated with the development of T1D. Here, we f PTPN6 observed expression of PTPN6 and PTPN1 in human islets and islets from non-obese f cytokines diabetic (NOD) mice. To clarify the role of these PTPs in β-cells/islets, we took advantage f inflammation Journal of Molecular Endocrinology of CRISPR/Cas9 technology and pharmacological approaches to inactivate both proteins. We identify PTPN6 as a negative regulator of TNF-α-induced β-cell death, through JNK- dependent BCL-2 protein degradation.
    [Show full text]
  • The Expression Patterns and the Prognostic Roles of PTPN Family Members in Digestive Tract Cancers
    Preprint: Please note that this article has not completed peer review. The expression patterns and the prognostic roles of PTPN family members in digestive tract cancers CURRENT STATUS: UNDER REVIEW Jing Chen The First Affiliated Hospital of China Medical University Xu Zhao Liaoning Vocational College of Medicine Yuan Yuan The First Affiliated Hospital of China Medical University Jing-jing Jing The First Affiliated Hospital of China Medical University [email protected] Author ORCiD: https://orcid.org/0000-0002-9807-8089 DOI: 10.21203/rs.3.rs-19689/v1 SUBJECT AREAS Cancer Biology KEYWORDS PTPN family members, digestive tract cancers, expression, prognosis, clinical features 1 Abstract Background Non-receptor protein tyrosine phosphatases (PTPNs) are a set of enzymes involved in the tyrosyl phosphorylation. The present study intended to clarify the associations between the expression patterns of PTPN family members and the prognosis of digestive tract cancers. Method Expression profiling of PTPN family genes in digestive tract cancers were analyzed through ONCOMINE and UALCAN. Gene ontology enrichment analysis was conducted using the DAVID database. The gene–gene interaction network was performed by GeneMANIA and the protein–protein interaction (PPI) network was built using STRING portal couple with Cytoscape. Data from The Cancer Genome Atlas (TCGA) were downloaded for validation and to explore the relationship of the PTPN expression with clinicopathological parameters and survival of digestive tract cancers. Results Most PTPN family members were associated with digestive tract cancers according to Oncomine, Ualcan and TCGA data. For esophageal carcinoma (ESCA), expression of PTPN1, PTPN4 and PTPN12 were upregulated; expression of PTPN20 was associated with poor prognosis.
    [Show full text]
  • The Regulatory Roles of Phosphatases in Cancer
    Oncogene (2014) 33, 939–953 & 2014 Macmillan Publishers Limited All rights reserved 0950-9232/14 www.nature.com/onc REVIEW The regulatory roles of phosphatases in cancer J Stebbing1, LC Lit1, H Zhang, RS Darrington, O Melaiu, B Rudraraju and G Giamas The relevance of potentially reversible post-translational modifications required for controlling cellular processes in cancer is one of the most thriving arenas of cellular and molecular biology. Any alteration in the balanced equilibrium between kinases and phosphatases may result in development and progression of various diseases, including different types of cancer, though phosphatases are relatively under-studied. Loss of phosphatases such as PTEN (phosphatase and tensin homologue deleted on chromosome 10), a known tumour suppressor, across tumour types lends credence to the development of phosphatidylinositol 3--kinase inhibitors alongside the use of phosphatase expression as a biomarker, though phase 3 trial data are lacking. In this review, we give an updated report on phosphatase dysregulation linked to organ-specific malignancies. Oncogene (2014) 33, 939–953; doi:10.1038/onc.2013.80; published online 18 March 2013 Keywords: cancer; phosphatases; solid tumours GASTROINTESTINAL MALIGNANCIES abs in sera were significantly associated with poor survival in Oesophageal cancer advanced ESCC, suggesting that they may have a clinical utility in Loss of PTEN (phosphatase and tensin homologue deleted on ESCC screening and diagnosis.5 chromosome 10) expression in oesophageal cancer is frequent, Cao et al.6 investigated the role of protein tyrosine phosphatase, among other gene alterations characterizing this disease. Zhou non-receptor type 12 (PTPN12) in ESCC and showed that PTPN12 et al.1 found that overexpression of PTEN suppresses growth and protein expression is higher in normal para-cancerous tissues than induces apoptosis in oesophageal cancer cell lines, through in 20 ESCC tissues.
    [Show full text]
  • Targeting Protein Tyrosine Phosphatases in Cancer Lakshmi Reddy Bollu, Abhijit Mazumdar, Michelle I
    Published OnlineFirst January 13, 2017; DOI: 10.1158/1078-0432.CCR-16-0934 Molecular Pathways Clinical Cancer Research Molecular Pathways: Targeting Protein Tyrosine Phosphatases in Cancer Lakshmi Reddy Bollu, Abhijit Mazumdar, Michelle I. Savage, and Powel H. Brown Abstract The aberrant activation of oncogenic signaling pathways is a act as tumor suppressor genes by terminating signal responses universal phenomenon in cancer and drives tumorigenesis and through the dephosphorylation of oncogenic kinases. More malignant transformation. This abnormal activation of signal- recently, it has become clear that several PTPs overexpressed ing pathways in cancer is due to the altered expression of in human cancers do not suppress tumor growth; instead, they protein kinases and phosphatases. In response to extracellular positively regulate signaling pathways and promote tumor signals, protein kinases activate downstream signaling path- development and progression. In this review, we discuss both ways through a series of protein phosphorylation events, ulti- types of PTPs: those that have tumor suppressor activities as mately producing a signal response. Protein tyrosine phospha- well as those that act as oncogenes. We also discuss the tases (PTP) are a family of enzymes that hydrolytically remove potential of PTP inhibitors for cancer therapy. Clin Cancer Res; phosphate groups from proteins. Initially, PTPs were shown to 23(9); 1–7. Ó2017 AACR. Background in cancer and discuss the current status of PTP inhibitors for cancer therapy. Signal transduction is a complex process that transmits extra- PTPs belong to a superfamily of enzymes that hydrolytically cellular signals effectively through a cascade of events involving remove phosphate groups from proteins (2).
    [Show full text]
  • Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance
    ORIGINAL RESEARCH published: 18 August 2020 doi: 10.3389/fimmu.2020.01856 Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance Larissa Hering 1, Egle Katkeviciute 1, Marlene Schwarzfischer 1, Philipp Busenhart 1, Claudia Gottier 1, Dunja Mrdjen 2, Juliana Komuczki 2†, Marcin Wawrzyniak 1, Silvia Lang 1, Kirstin Atrott 1, Burkhard Becher 2, Gerhard Rogler 1,3, Michael Scharl 1,3* and Marianne R. Spalinger 1 1 Department of Gastroenterology and Hepatology, University Hospital Zurich, University of Zurich, Zurich, Switzerland, 2 Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland, 3 Zurich Center for Integrative Human Physiology, University of Zurich, Zurich, Switzerland Edited by: Michele Marie Kosiewicz, University of Louisville, United States Protein tyrosine phosphatase non-receptor type 2 (PTPN2) plays a pivotal role in Reviewed by: immune homeostasis and has been associated with human autoimmune and chronic Cong-Yi Wang, inflammatory diseases. Though PTPN2 is well-characterized in lymphocytes, little is Tongji Medical College, China Andrew L. Mellor, known about its function in innate immune cells. Our findings demonstrate that dendritic Newcastle University, United Kingdom cell (DC)-intrinsic PTPN2 might be the key to explain the central role for PTPN2 in *Correspondence: the immune system to maintain immune tolerance. Partial genetic PTPN2 ablation in Michael Scharl DCs resulted in spontaneous inflammation, particularly in skin, liver, lung and kidney 22 [email protected] weeks post-birth. DC-specific PTPN2 controls steady-state immune cell composition † Present address: and even incomplete PTPN2 deficiency in DCs resulted in enhanced organ infiltration of Juliana Komuczki, Roche Innovation Center Basel, conventional type 2 DCs, accompanied by expansion of IFNγ-producing effector T-cells.
    [Show full text]
  • The Role of Protein Tyrosine Phosphatases in Inflammasome
    International Journal of Molecular Sciences Review The Role of Protein Tyrosine Phosphatases in Inflammasome Activation Marianne R. Spalinger 1,* , Marlene Schwarzfischer 1 and Michael Scharl 1,2 1 Department of Gastroenterology and Hepatology, University Hospital Zurich, 8091 Zurich, Switzerland; Marlene.Schwarzfi[email protected] (M.S.); [email protected] (M.S.) 2 Zurich Center for Integrative Human Physiology, University of Zurich, 8006 Zurich, Switzerland * Correspondence: [email protected]; Tel.: +41-44-255-3794 Received: 16 July 2020; Accepted: 29 July 2020; Published: 31 July 2020 Abstract: Inflammasomes are multi-protein complexes that mediate the activation and secretion of the inflammatory cytokines IL-1β and IL-18. More than half a decade ago, it has been shown that the inflammasome adaptor molecule, ASC requires tyrosine phosphorylation to allow effective inflammasome assembly and sustained IL-1β/IL-18 release. This finding provided evidence that the tyrosine phosphorylation status of inflammasome components affects inflammasome assembly and that inflammasomes are subjected to regulation via kinases and phosphatases. In the subsequent years, it was reported that activation of the inflammasome receptor molecule, NLRP3, is modulated via tyrosine phosphorylation as well, and that NLRP3 de-phosphorylation at specific tyrosine residues was required for inflammasome assembly and sustained IL-1β/IL-18 release. These findings demonstrated the importance of tyrosine phosphorylation as a key modulator of inflammasome activity. Following these initial reports, additional work elucidated that the activity of several inflammasome components is dictated via their phosphorylation status. Particularly, the action of specific tyrosine kinases and phosphatases are of critical importance for the regulation of inflammasome assembly and activity.
    [Show full text]
  • Recombinant PTPN2 Protein Catalog No: 31592, 31992 Quantity: 50, 1000 Μg Expressed In: E
    Recombinant PTPN2 protein Catalog No: 31592, 31992 Quantity: 50, 1000 µg Expressed In: E. coli Concentration: 0.5 µg/µl Source: Human Buffer Contents: Recombinant PTPN2 protein is supplied at a concentration of 0.5 µg/µl in 25 mM Tris pH 8.0, 300 mM NaCl, 5% glycerol. Background: PTPN2 (Protein Tyrosine Phosphatase, Non-Receptor Type 2, also known as TCELLPTP, TCPTP, TC- PTP, PTPT, PTN2) is a member of the protein tyrosine phosphatase (PTP) family. Members of the PTP family have a highly conserved catalytic motif, which is essential for the catalytic activity. PTPs regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation as signaling molecules. Epidermal growth factor receptor (EGFR) and the adaptor protein Shc were reported to be substrates of this PTP, which suggested the roles in growth factor mediated cell signaling. PTPN2 dephosphorylates multiple substrates including receptor protein tyrosine kinases INSR, EGFR, CSF1R, PDGFR and non-receptor protein tyrosine kinases Recombinant PTPN2 protein gel. like JAK1, JAK2, JAK3, Src family kinases, STAT1, STAT3, STAT5A, STAT5B Recombinant PTPN2 protein was run and STAT6 either in the nucleus or the cytoplasm. on a 10% SDS-PAGE gel and stained with Coomassie Blue. Protein Details: Recombinant PTPN2 was expressed in E. coli cells as the full MW: 75.7 kDa length protein (accession number NP_002819.2) with an N-terminal GST tag. The molecular weight of the protein is 75.7 kDa. Purity: > 50% Application Notes: This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Activity Assay Conditions: 1 mM pNPP was incubated with different concentrations of PTPN2 protein in reaction buffer including 50 mM HEPES pH 7.5, 2 mM EDTA, 3 mM DTT, 100 mM NaCl at room temperature.
    [Show full text]
  • Lipid Phosphatases Identified by Screening a Mouse Phosphatase Shrna Library Regulate T-Cell Differentiation and Protein Kinase
    Lipid phosphatases identified by screening a mouse PNAS PLUS phosphatase shRNA library regulate T-cell differentiation and Protein kinase B AKT signaling Liying Guoa, Craig Martensb, Daniel Brunob, Stephen F. Porcellab, Hidehiro Yamanea, Stephane M. Caucheteuxa, Jinfang Zhuc, and William E. Paula,1 aCytokine Biology Unit, cMolecular and Cellular Immunoregulation Unit, Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and bGenomics Unit, Research Technologies Section, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840 Contributed by William E. Paul, March 27, 2013 (sent for review December 18, 2012) Screening a complete mouse phosphatase lentiviral shRNA library production (10, 11). Conversely, constitutive expression of active using high-throughput sequencing revealed several phosphatases AKT leads to increased proliferation and enhanced Th1/Th2 cy- that regulate CD4 T-cell differentiation. We concentrated on two lipid tokine production (12). phosphatases, the myotubularin-related protein (MTMR)9 and -7. The amount of PI[3,4,5]P3 and the level of AKT activation are Silencing MTMR9 by shRNA or siRNA resulted in enhanced T-helper tightly controlled by several mechanisms, including breakdown of (Th)1 differentiation and increased Th1 protein kinase B (PKB)/AKT PI[3,4,5]P3, down-regulation of the amount and activity of PI3K, phosphorylation while silencing MTMR7 caused increased Th2 and and the dephosphorylation of AKT (13). PTEN is a major negative Th17 differentiation and increased AKT phosphorylation in these regulator of PI[3,4,5]P3. It removes the 3-phosphate from the cells.
    [Show full text]
  • PTPN2 Expression Is Associated with Inferior Molecular Response in De-Novo Chronic Myeloid Leukaemia Patients
    Letters to the Editor 702 CONFLICT OF INTEREST 4 Wang Y, Krivtsov AV, Sinha AU, North TE, Goessling W, Feng Z et al. The authors declare no conflict of interest. The Wnt/beta-catenin pathway is required for the development of leukemia stem cells in AML. Science 2010; 327: 1650–1653. 5 Yeung J, Esposito MT, Gandillet A, Zeisig BB, Griessinger E, Bonnet D et al. beta- ACKNOWLEDGEMENTS Catenin mediates the establishment and drug resistance of MLL leukemic stem cells. Cancer Cell 2010; 18: 606–618. This work was supported in part by the National Cancer Institute (CA66996 and 6 Zhao C, Blum J, Chen A, Kwon HY, Jung SH, Cook JM et al. Loss of beta-catenin CA140575) and the Leukemia and Lymphoma Society. DK was supported by NIH impairs the renewal of normal and CML stem cells in vivo. Cancer Cell 2007; 12: NIDDK award K01DK092300. 528–541. 7 Cobas M, Wilson A, Ernst B, Mancini SJ, MacDonald HR, Kemler R et al. 1 2 2 1 2 CEL Ng , A Sinha , A Krivtsov , S Dias , J Chang , Beta-catenin is dispensable for hematopoiesis and lymphopoiesis. J Exp Med 2004; SA Armstrong1,2 and D Kalaitzidis1,3 199: 221–229. 1Division of Hematology/Oncology, Children’s Hospital Boston and 8 Ward AF, Braun BS, Shannon KM. Targeting oncogenic Ras signaling in hemato- Dana-Farber Cancer Institute, Harvard Medical School and the logic malignancies. Blood 2012; 120: 3397–3406. Harvard Stem Cell Institute, Boston, MA 02115, USA 9 Braun BS, Tuveson DA, Kong N, Le DT, Kogan SC, Rozmus J et al.
    [Show full text]
  • Mapping the Human Phosphatome on Growth Pathways
    Molecular Systems Biology 8; Article number 603; doi:10.1038/msb.2012.36 Citation: Molecular Systems Biology 8:603 & 2012 EMBO and Macmillan Publishers Limited All rights reserved 1744-4292/12 www.molecularsystemsbiology.com Mapping the human phosphatome on growth pathways Francesca Sacco1,*, Pier Federico Gherardini1, Serena Paoluzi1, Julio Saez-Rodriguez2, Manuela Helmer-Citterich1, Antonella Ragnini-Wilson1,3, Luisa Castagnoli1 and Gianni Cesareni1,4,* 1 Department of Biology, University of Rome ‘Tor Vergata’, Rome, Italy, 2 EMBL-EBI, Hinxton, UK, 3 High-throughput Microscopy Facility, Department of Translational and Cellular Pharmacology, Consorzio Mario Negri Sud, SM Imbaro, Italy and 4 IRCCS Fondazione Santa Lucia, Rome, Italy * Corresponding authors. F Sacco or G Cesareni, Department of Biology, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica, Rome 00133, Italy. Tel.: þ 39 0672594315; Fax: þ 39 06202350; E-mail: [email protected] or [email protected] Received 5.3.12; accepted 10.7.12 Large-scale siRNA screenings allow linking the function of poorly characterized genes to phenotypic readouts. According to this strategy, genes are associated with a function of interest if the alteration of their expression perturbs the phenotypic readouts. However, given the intricacy of the cell regulatory network, the mapping procedure is low resolution and the resulting models provide little mechanistic insights. We have developed a new strategy that combines multi- parametric analysis of cell perturbation with logic modeling to achieve a more detailed functional mapping of human genes onto complex pathways. A literature-derived optimized model is used to infer the cell activation state following upregulation or downregulation of the model entities.
    [Show full text]