1. Manifestation of Novel Social Challenges of the European Union in the Teaching Material of Medical Biotechnology Master’s Programmes at the University of Pécs and at the University of Debrecen Identification number: TÁMOP-4.1.2-08/1/A-2009-0011

2. TYROSINE KINASE- LINKED RECEPTORS PART 1 CYTOKINE-CHEMOKINE SIGNALING Tímea Berki and Ferenc Boldizsár Signal transduction Manifestation of Novel Social Challenges of the European Union in the Teaching Material of Medical Biotechnology Master’s Programmes at the University of Pécs and at the University of Debrecen Identification number: TÁMOP-4.1.2-08/1/A-2009-0011

3. TÁMOP-4.1.2-08/1/A-2009-0011 Cytokine groupsStructural: 1 14 a-helix bundle family IL-2 subfamily IFN subfamily IL-10 subfamily 2 2IL-1 family 3 3IL-17 familyFunctional: 1 1Haematopoietic (EPO, TPO, G-CSF, GM-CSF, SCF etc.) 2 2Lymphocyte differentiation/acti vation Th1: IL-2, TNF, IFN  Th2: IL-4, IL-5, Il- 13; IL-17, IL-23 3 3Inflammatory (IL-1, IL-6, TNF  )

4. TÁMOP-4.1.2-08/1/A-2009-0011 Cytokine receptors Interferon- , - β, and -  receptor, IL-10 receptor Tumor necrosis factor (TNF) receptors I and II, CD40, Fas (Apo1, CD95), CD30, CD27, nerve growth factor receptor CCR1-10, CXCR1-5, XCR1, CX3CR1 Receptor for IL-2, IL-4, IL-7, IL-9 and IL-15 share a common chain CD132 or  c (common gamma chain). Il-2 receptor also has a third chain, a high affinity subunit IL-2R  (CD25) Receptors for erythropoietin, growth hormone, and IL-13 Receptor for IL-3, IL-5, and GM-CSF share a common chain,CD131 or β c (common beta chain) Class I cytokine receptor (hematopoietin receptor family) Class II cytokine receptor TNF-receptor family Chemokine-receptor family  β

5. TÁMOP-4.1.2-08/1/A-2009-0011 Characteristics of multichain cytokine receptors gp130 LIF/OSM IL-6 gp130 IL-11 gp130 CNTF CNTFR gp130 GM-CSFR  GM-CSF β IL-3R IL-3 β IL-5R IL-5 β IL-2R  IL-2R β IL-2  IL-2R β IL-15R  IL-15  IL-7R IL-7  IL-9R IL-9  IL-4R IL-4  GM-CSF receptor subfamily (common β subunit) Il-6 receptor subfamily (common gp130 subunit) Il-2 receptor subfamily (common  subunit)

6. TÁMOP-4.1.2-08/1/A-2009-0011 Overview of cytokine signalling Cytokine receptors consist of at least two chains, the cytoplasmatic domains of which bind Janus kinases (JAKs) JAK Transcription factors (STATs) bind to the phosphorylated receptors, and are in turn phosphorylated by the activated JAKs Phosphorylated STATs form dimers that translocate into the nucleus to initiate new gene transcription JAK PP PP PP Cytokine binding dimerizes the receptor, bringing together the cytoplasmic JAKs, which activate each other and phosphorylate the receptor JAK PP PP PP P P P P STAT

7. TÁMOP-4.1.2-08/1/A-2009-0011 The structure of JAK and STAT proteins FERMyKIKI"SH 2 " JH7JH6JH5JH4JH3JH2 JH1 020040060080010001200 NH 2 Coiled coilDBDLkSH 2 TAD Y P 0200400600800 JAK structure STAT structure

8. TÁMOP-4.1.2-08/1/A-2009-0011 I JAK I FERMyKIKI"SH 2 " JH7JH6JH5JH4JH3JH2JH1 020040060080010001200 Kinase Pseudokinase FERM domain: band 4.1, ezrin, radixin and moesin FERM domain: band 4.1, ezrin, radixin and moesin JH= Janus homology domain Proline rich membrane proximal part of Cytokine Receptors

9. TÁMOP-4.1.2-08/1/A-2009-0011 II JAK II JAK 1 IFN-I/II, IL-2,-4,- 6,-10 JAK2 IFN-II, IL-3,-5, GM-CSF JAK3 IL-2 ,-4,-7,-9,- 15,-21 TYK2 IFN-I, IL-6,-10,- 12,-23

10. TÁMOP-4.1.2-08/1/A-2009-0011 I STAT I NH 2 Coiled coilDBDLkSH 2 TAD Y P 0200400600800 DNA-binding Transcriptional activation Linker Receptor recruitment and dimerization Binding of regulators Dimerization DNA-binding Nuclear transport

11. TÁMOP-4.1.2-08/1/A-2009-0011 Human STATs NameChr (Hu)MW (kDa)FunctionUpstream STAT12q32.291 IFN-  /IL-12 axis (Th1 diff.) Y 701 EGFR, FGFR3/4, JAK1/2, Lck, KIT, TYK2 STAT212q13.3113Y 690 JAK1 S 727 PKCd, CamKIIg, Ribosomal protein S6 kinase alpha 5 STAT317q21.31Th17 Y 705 FGFR3/4, Hck, JAK1/2, cSrc, EphA3 S 727 ERK1/2, PKCd, MAPK8, Ribosomal protein S6 kinase alpha 5 STAT4 2q32.2-q32.3 Th1S 721 MAPK14, MAP2K6 STAT5A17q11.2Y 694 JAK2, Btk STAT5B17q11.2 Y 679 cSrc Y 699, 725, 740, 743 EGFR STAT612q13IL-4 (Th2 diff.)JAK1/2/3

12. TÁMOP-4.1.2-08/1/A-2009-0011 STAT IISTAT1IFN-I/II Inflammatio n STAT2IFN-I STAT3 IL-6 and IL-10 families, IL-21, IL-27 STAT4IL-12,-23Th1, Th17 STAT5A and B IL-3,-5, GM-CSF STAT6IL-4,-13 Th2, allergy

13. TÁMOP-4.1.2-08/1/A-2009-0011 Regulation of JAK/STAT signaling Phosphatases SHP-1/2, CD45 – JAK SHP-2, PTP1B, TC-PTP, PTP-BL – STAT Nuclear export/import NES (nuclear export sequence) NLS (nuclear localization sequence) SOCS (suppressors of cytokine signaling) eg. PIAS (Protein Inhibitor of Activated STATs) Ser-phosphorylation, acetylation, o- glycosylation of TAD

14. TÁMOP-4.1.2-08/1/A-2009-0011 JAK inhibitorsTreatment: Polycythemia vera, thrombocytemia, Myeloid metaplasia, myelofibrosis Psoriasis RAExamples: Lestaurtinib Tofacitinib Ruxolitinib

15. TÁMOP-4.1.2-08/1/A-2009-0011 Chemokines 90-130 aa. polypeptidesFunctions: Chemotaxis for different leukocytes: –Regulation of normal leukocyte traffic –Recruitment of cells to inflammatory sites Enhancement of cell adhesion Activation of effector leukocytes Development of the inflammatory reaction Development of normal lymphoid tissues

16. TÁMOP-4.1.2-08/1/A-2009-0011 Chemokine signal through receptors coupled with G- proteins Chemokine receptorCa 2+ channels G protein cAMPcAMP Adenylyl cyclase Adenylyl cyclase Adhesion Chemotaxis ? Differentiation, proliferation Ras ? Actin polymerization PLC β 2 DAG IP 3 Ca 2+ PKC Cytoskeletal rearrangment ? GDP  β  GTP   β

17. TÁMOP-4.1.2-08/1/A-2009-0011 Chemokine signaling pathways Chemokine receptor Ca 2+ channel Gene expression and apoptosis Chemokines Ca 2+ RAS DAG JNK p38 Ras pathway Ras pathway  β GRB2 NF-  B STAT5 GTP  Elk- 1 MAPK pathway MAPK pathway PTK Cell activation PLA2 Rho pathway Rho pathway Rho SRF InternalizationDegradationRecyclingInternalizationDegradationRecycling GRK β -Arrestin PLC β PKC β Akt/PKB pathway Akt/PKB pathway Akt/ PKB Akt/ PKB PDK NF-  B Elk- 1 Plasma membrane Cytoplasm Nucleus PIP 2 IP 3 GDP  β  JAK2