Cell Adhesion and Signaling

Transcription

1 Cell Adhesion and Signaling Institute of Anatomy and Cell Biology 1

2 Transactivation NATURE REVIEWS CANCER VOLUME 7 FEBRUARY

3 Functions of Cell Adhesion cell cycle proliferation migration differentiation survival Cell Adhesion inflammation cell shape angiogenesis polarity apoptosis 3

4 CELL-CELL ADHESION Cadherins E, N, P cadherins desmosomal cadherins Ca2+- or Mg2+ -dependence homophilic actin filaments (via catenins) yes homophilic intermediate filaments (via desmoplakins, plakoglobin and other proteins) Ig family members N-CAM, L1 no homophilic or heterophilic Selectins (blood cells + endothelial cells) Integrins (on blood cells) LFA-1 (a L b 2 ), Mac-1 (a M b 2 ) unknown P-selectin yes heterophilic unknown no yes heterophilic actin filaments no adhesion belts desmosomes no CELL-MATRIX ADHESION Integrins many types yes heterophilic actin filaments (via talin, vinculin, and other proteins) a 6 b 4 yes heterophilic intermediate filaments focal contacts hemidesmosomes Transmembrane Proteoglycans syndecans no heterophilic actin filaments no 4

5 Major families of cell-adhesion molecules 5

6 Annu. Rev. Pharmacol. Toxicol :

7 Annu. Rev. Pharmacol. Toxicol :

8 Significance of cytoskeletons in cell adhesion without cytoskeletal attachment adhesion molecule Plasma membrane No effect on cell PULL cytoskeleton with cytoskeletal attachment Cell adhesion PULL 8

9 Selectins Chemoattractants Integrins Ig CAMs 1. Tethering and Rolling 2. Activation 3. Firm adhesion 4. Transmigration endothelium Inflammatory stimulus 9 9

10 Selectin P-selectin Lectin domain E-selectin EGF-like domain L-selectin Short consensus repeat 10

11 LEUKOCYTE L-Selectin P-Selectin E-Selectin ENDOTHELIAL CELLS Thrombin Histamine TNF- α LPS TNF- α IL-1 β IFN- γ LPS 11

12 L-Selectin ESL-1 PSGL-1 LEUKOCYTE h ENDOTHELIAL CELL GlyCAM-1 CD34 MAdCAM-1 E-Selectin P-Selectin 12

13 INTERACTION Selectin Selectin Ligand Protein Carbohydrate on lipids proteins 13

14 Summary of intracellular signaling cascades in leukocytes on L-selectin triggering P-Ser PKC p56lck (J. Biol. Chem., V. 279, , August 13, 2004) cell growth, survival, apoptosis 14 Adapted from Pediatric Research 48: (2000)

15 Molecular Events in Transmembrane Signaling via E-selectin E-selectin (HUVEC cell) P-Tyr603- -Tyr-P SHP2 Shc Grb2 Sos Ras Raf MEK ERK1/2 J. Biol. Chem., Vol. 276, Issue 51, , December 21, 2001 gene expression (ex: c-fos) 15

16 Regulation of transendothelial migration of colon cancer cells by E-selectin-mediated activation of p38 and ERK MAP kinases Tremblay PL, Auger FA, Huot J. Oncogene Oct 26;25(50):

17 Signaling cascades initiated by ligation of either the selectins or their ligands Calcium flux Src family kinases Ras activation MAP Kinases Neutral sphingomyelinase β2 integrin activation Cell shape change Oxygen radical generation Gene expression (ex: TNFα, IL-8, CXCR4) 17 Adapted from seminars in IMMUNOLOGY, Vol. 14, 2002: pp

18 Integrin Cell, Vol. 110, , September 20,

19 Signaling transduction pathways emanating from Integrins 31 MAY 2002 VOL 296 p SCIENCE 19

20 NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 2 NOVEMBER

21 Common integrins and ECM ligands bound 21 Journal of Cell Science 115,

22 Integrin binding proteins 22 Annu. Rev. Pharmacol. Toxicol :

23 Integrin binding proteins 23 Annu. Rev. Pharmacol. Toxicol :

24 Integrin Signaling Pathway Integrins 24 May 31, J. Thomas Parsons

25 FAK Focal Adhesion Kinase 25

26 Oncogene (2004) 23,

27 Focal Adhesion Kinase (FAK) nonreceptor tyrosine kinase NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 6 JANUARY

28 Example of FAK function caveolin Focal Adhesion Kinase P P Shc Ras JNK CrkII C3G Ras survival Rap-1 B-Raf Raf ERK contractibility ERK 28 Modified from Annu. Rev. Pharmacol. Toxicol :

29 Multiple signaling proteins dock at Tyrosine 397 of FAK py Biochimica et Biophysica Acta 1540 (2001) 1-21

30 NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 6 JANUARY

31 Scanning electron micrograph of a basal lamina in the cornea of a chick embryo Some of the epithelial cells (E) have been removed to expose the upper surface of the matlike basal lamina (BL). A network of collagen fibrils (C) in the underlying connective tissue interacts with the lower face of the lamina. (Courtesy of Robert Trelstad.) 31

32 a Mock 4GalNAc-T3 c b d Mock 4GalNAc-T3 32

33 a Mock 4GalNAc-T3 0 h 32 h Migration rate ( m/h) 10 * Mock 4GalNAc-T3 33

34 b Mock 4GalNAc-T3 Mock 4GalNAc-T3 34

35 A C Mock 4GalNAc-T IP: FAK (C20) Mock 4GalNAc-T3 IB: 4GalNAc-T3 4G10 (anti-py) PY20 (anti-py) FAK IB: B D Mock 4GalNAc-T IB: py397 py576 FAK IP: FAK (C20) Mock 4GalNAc-T3 perk1/ py118-paxillin py31-paxillin ERK1/ paxillin WCL -actin 35

36 ILK Integrin-Linked Kinase 36

37 Integrin-Linked Kinase binding proteins The Journal of Cell Biology, Volume 155, Number 4, November 12,

38 ILK-PINCH-parvin (IPP) complex NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 7 JANUARY

39 Signaling through IPP complex NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 7 JANUARY

40 NATURE REVIEWS CANCER VOLUME 5 JANUARY

41 Activation of MAP kinase cascades by integrins 41

42 Activation of MAP kinase cascades by integrins caveolin Focal Adhesion Kinase Shc P P Ras JNK CrkII C3G Ras survival Rap-1 B-Raf Raf ERK contractibility ERK 42 Modified from Annu. Rev. Pharmacol. Toxicol :

43 Integrin regulates actin filaments formation via Rho GTPases (Rho, Rac, CDC42) 43

44 Integrin signaling to the cytoskeleton via Rho GTPases Rho Rac CDC42 FILOPODIA STRESS FIBERS P cofilin Lamellipodia 44 Annu. Rev. Pharmacol. Toxicol :

45 Mechanism of activation of N-WASP P Active N-WASP Adapted from Journal of Cell Science 114, ,

46 Mechanism of activation of WAVE2 Journal of Cell Science 114, ,

47 Cooperativity between growth-factor receptors and integrins integrin growth factor receptor 47 NATURE CELL BIOLOGY VOL 2 DECEMBER 2000 E

48 Integrin regulates signaling in RTK/Ras/MAPK pathway integrin Receptor Tyrosine Kinase 2 1 cytoplasm 3 nucleus 48 Annu. Rev. Pharmacol. Toxicol :

49 1. Activation of receptor tyrosine kinase (RTK) Integrin aggregation and ligand binding EGFR, PDGF-R, FGFR phosphorylation Integrin aggregation alone PDGFβ-R, EGFR Direct integrin-rtk interaction αvβ3 and PDGFβ-R, insulin receptor, VEGF-R2 Via integrin-associated cytoskeletal components Integrin-actin-Neu RTK, integrin-fak- EGFR or PDGFR 49

50 Integrin regulation of growth factor receptors NATURE CELL BIOLOGY VOL4 APRIL 2002 E

51 FAK integrates growth-factor and integrin signals to promote cell migration Current Opinion in Genetics & Development 2004, 14:

52 2. Coupling between upstream and downstream molecules Between Ras-Raf EGFR, PDGF-R in anchorage-dependent cell growth 52

53 3. Transmission of the signal from cytoplasm to nucleus J Cell Biol 2001 Apr 16;153(2): Integrin-mediated adhesion Regulates ERK Nuclear Translocation and Phosphorylation of Elk-1 Actin cytoskeletondependent (NIH 3T3 cells) G1 phase progression 53

54 Examples of synergisms between integrins and RTK signaling 54 Annu. Rev. Pharmacol. Toxicol :