Cell Adhesion and Signaling mchuang@ntu.edu.tw Institute of Anatomy and Cell Biology 1
Transactivation NATURE REVIEWS CANCER VOLUME 7 FEBRUARY 2007 85 2
Functions of Cell Adhesion cell cycle proliferation migration differentiation survival Cell Adhesion inflammation cell shape angiogenesis polarity apoptosis 3
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
Major families of cell-adhesion molecules 5
Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323 6
Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323 7
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
Selectins Chemoattractants Integrins Ig CAMs 1. Tethering and Rolling 2. Activation 3. Firm adhesion 4. Transmigration endothelium Inflammatory stimulus 9 9
Selectin P-selectin Lectin domain E-selectin EGF-like domain L-selectin Short consensus repeat 10
LEUKOCYTE L-Selectin P-Selectin E-Selectin ENDOTHELIAL CELLS Thrombin Histamine TNF- α LPS TNF- α IL-1 β IFN- γ LPS 11
L-Selectin ESL-1 PSGL-1 LEUKOCYTE h ENDOTHELIAL CELL GlyCAM-1 CD34 MAdCAM-1 E-Selectin P-Selectin 12
INTERACTION Selectin Selectin Ligand Protein Carbohydrate on lipids proteins 13
Summary of intracellular signaling cascades in leukocytes on L-selectin triggering P-Ser PKC p56lck (J. Biol. Chem., V. 279, 34472-34480, August 13, 2004) cell growth, survival, apoptosis 14 Adapted from Pediatric Research 48:283-288 (2000)
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, 48549-48553, December 21, 2001 gene expression (ex: c-fos) 15
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. 2006 Oct 26;25(50):6563-73. 16
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. 73 81
Integrin Cell, Vol. 110, 673 687, September 20, 2002 18
Signaling transduction pathways emanating from Integrins 31 MAY 2002 VOL 296 p.1652-1653 SCIENCE 19
NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 2 NOVEMBER 2001 795 20
Common integrins and ECM ligands bound 21 Journal of Cell Science 115, 3729-3738 2002
Integrin binding proteins 22 Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323
Integrin binding proteins 23 Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323
Integrin Signaling Pathway Integrins 24 http://stke.sciencemag.org May 31, 2002. J. Thomas Parsons
FAK Focal Adhesion Kinase 25
Oncogene (2004) 23, 7928 7946. 26
Focal Adhesion Kinase (FAK) nonreceptor tyrosine kinase NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 6 JANUARY 2005 59 27
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. 2002. 42:283 323
Multiple signaling proteins dock at Tyrosine 397 of FAK py397 29 Biochimica et Biophysica Acta 1540 (2001) 1-21
NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 6 JANUARY 2005 58 30
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
a Mock 4GalNAc-T3 c b d Mock 4GalNAc-T3 32
a Mock 4GalNAc-T3 0 h 32 h Migration rate ( m/h) 10 * 8 6 4 2 0 Mock 4GalNAc-T3 33
b Mock 4GalNAc-T3 Mock 4GalNAc-T3 34
A C Mock 4GalNAc-T3 1.0 2.5 1.0 2.7 1.0 3.5 IP: FAK (C20) Mock 4GalNAc-T3 IB: 4GalNAc-T3 4G10 (anti-py) PY20 (anti-py) FAK IB: B D Mock 4GalNAc-T3 1.0 2.5 1.0 1.7 IB: py397 py576 FAK IP: FAK (C20) Mock 4GalNAc-T3 perk1/2 1.0 1.8 py118-paxillin py31-paxillin 1.0 1.8 ERK1/2 1.0 0.5 paxillin WCL -actin 35
ILK Integrin-Linked Kinase 36
Integrin-Linked Kinase binding proteins The Journal of Cell Biology, Volume 155, Number 4, November 12, 2001 505-510 37
ILK-PINCH-parvin (IPP) complex NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 7 JANUARY 2006 22 38
Signaling through IPP complex NATURE REVIEWS MOLECULAR CELL BIOLOGY VOLUME 7 JANUARY 2006 24 39
NATURE REVIEWS CANCER VOLUME 5 JANUARY 2005 60 40
Activation of MAP kinase cascades by integrins 41
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. 2002. 42:283 323
Integrin regulates actin filaments formation via Rho GTPases (Rho, Rac, CDC42) 43
Integrin signaling to the cytoskeleton via Rho GTPases Rho Rac CDC42 FILOPODIA STRESS FIBERS P cofilin Lamellipodia 44 Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323
Mechanism of activation of N-WASP P Active N-WASP Adapted from Journal of Cell Science 114, 1801-1809, 2001 45
Mechanism of activation of WAVE2 Journal of Cell Science 114, 1801-1809, 2001 46
Cooperativity between growth-factor receptors and integrins integrin growth factor receptor 47 NATURE CELL BIOLOGY VOL 2 DECEMBER 2000 E225-229
Integrin regulates signaling in RTK/Ras/MAPK pathway integrin Receptor Tyrosine Kinase 2 1 cytoplasm 3 nucleus 48 Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323
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
Integrin regulation of growth factor receptors NATURE CELL BIOLOGY VOL4 APRIL 2002 E75-76 50
FAK integrates growth-factor and integrin signals to promote cell migration Current Opinion in Genetics & Development 2004, 14:92 101 51
2. Coupling between upstream and downstream molecules Between Ras-Raf EGFR, PDGF-R in anchorage-dependent cell growth 52
3. Transmission of the signal from cytoplasm to nucleus J Cell Biol 2001 Apr 16;153(2):273-82 Integrin-mediated adhesion Regulates ERK Nuclear Translocation and Phosphorylation of Elk-1 Actin cytoskeletondependent (NIH 3T3 cells) G1 phase progression 53
Examples of synergisms between integrins and RTK signaling 54 Annu. Rev. Pharmacol. Toxicol. 2002. 42:283 323