Cell Adhesion and Signaling

黃敏銓mchuang@ntu.edu.tw

Institute of Anatomy and Cell Biology 1

2

Transactivation

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Functions of Cell Adhesion

proliferation

differentiation

inflammation

angiogenesis

apoptosis

polarity

cell shape

survival

migration

cell cycle

Cell Adhesion

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4

CELL-CELL ADHESIONCadherins E, N, P

cadherinsCa2+- or Mg2+ -dependence

homophilic actin filaments (via catenins)

adhesion belts

desmosomalcadherins

yes homophilic intermediate filaments (via desmoplakins, plakoglobin and other proteins)

desmosomes

Ig family members N-CAM, L1 no homophilic or heterophilic

unknown no

Selectins (blood cells + endothelial cells)

P-selectin yes heterophilic unknown no

Integrins (on blood cells) LFA-1 (aLb2), Mac-1 (aMb2)

yes heterophilic actin filaments no

CELL-MATRIX ADHESIONIntegrins many types yes heterophilic actin filaments

(via talin, vinculin, and other proteins)

focal contacts

a6b4 yes heterophilic intermediate filaments

hemidesmosomes

TransmembraneProteoglycans

syndecans no heterophilic actin filaments no

Major families of cell-adhesion molecules

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Significance of cytoskeletons in cell adhesion

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No effect on cell

Cell adhesion

withoutcytoskeletalattachment

withcytoskeletalattachment

PULL

PULL

cytoskeleton

adhesion moleculePlasma membrane

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1. Tetheringand Rolling

3. Firm adhesion

4. Transmigration

Inflammatorystimulus

2. Activation

Selectins

Chemoattractants

Integrins

endothelium

Ig CAMs

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Selectin

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Short consensus repeat

EGF-like domain

Lectin domain

L-selectin

E-selectin

P-selectin

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L-Selectin

LEUKOCYTE

P-Selectin E-Selectin

ENDOTHELIAL CELLS

ThrombinHistamineTNF- αLPS

TNF- αIL-1 βIFN- γLPS

ESL-1 PSGL-1L-Selectin

LEUKOCYTE

h

ENDOTHELIAL CELL

GlyCAM-1 CD34 MAdCAM-1 E-Selectin P-Selectin

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Protein Carbohydrate

INTERACTION

Selectin Selectin Ligand

on lipids

proteins

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Summary of intracellular signaling cascades in leukocytes on L-selectin triggering

p56Lck

cell growth, survival, apoptosis

P-Ser

(J. Biol. Chem., V. 279, 34472-34480, August 13, 2004)

PKC

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Adapted from Pediatric Research 48:283-288 (2000)

Molecular Events in Transmembrane Signaling via E-selectin

E-selectin

SHP2 Shc

Grb2

Sos

Ras Raf MEK

ERK1/2

(HUVEC cell) -Tyr-PP-Tyr603-

15gene expression

(ex: c-fos)J. Biol. Chem., Vol. 276, Issue 51, 48549-48553, December 21, 2001

Regulation of transendothelial migration of colon cancer cells by E-selectin-mediated activation of p38 and ERK MAP kinases

Tremblay PL Auger FA

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, , Huot J. Oncogene. 2006 Oct 26;25(50):6563-73.

Signaling cascades initiated by ligation of either the selectins or their ligands

Calcium fluxSrc family kinasesRas activationMAP KinasesNeutral sphingomyelinase

Gene expression(ex: TNFα, IL-8, CXCR4)

Oxygen radicalgenerationCell shape changeβ2 integrin

activation

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Adapted from seminars in IMMUNOLOGY, Vol. 14, 2002: pp. 73–81

Integrin

18Cell, Vol. 110, 673–687, September 20, 2002

Signaling transduction pathways emanating from Integrins

31 MAY 2002 VOL 296 p.1652-1653 SCIENCE

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Common integrins and ECM ligands bound

Journal of Cell Science 115, 3729-3738 © 2002

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Integrin binding proteins

Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

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Integrin binding proteins

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Integrin Signaling Pathway

Integrins

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http://stke.sciencemag.org May 31, 2002. J. Thomas Parsons

FAKFocal Adhesion Kinase

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Oncogene (2004) 23, 7928–7946.

Focal Adhesion Kinase (FAK)nonreceptor tyrosine kinase

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Modified from Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Example of FAK function

Focal Adhesion Kinase

ERKERK

JNK

CrkII

C3G

Rap-1

B-Raf

Raf

Shc

survival

contractibility

P P

caveolin

RasRas

Multiple signaling proteins dock at Tyrosine 397 of FAK

Biochimica et Biophysica Acta 1540 (2001) 1-21

pY397

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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 cMock β4GalNAc-T3

Mock β4GalNAc-T3

b d

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aMock β4GalNAc-T3

0 h

32 h

Mock β4GalNAc-T302468

10 *

Mig

ratio

n ra

te (μ

m/h

)

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bMock β4GalNAc-T3

Mock β4GalNAc-T3

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BMock

IP: FAK (C20)

4G10(anti-pY)

FAK

IB:β4GalNAc-T3A

1.0 2.7

β4GalNAc-T3 IB:Mockβ4GalNAc-T3

1.0 2.5 1.0 2.5

1.0 1.7

pY397

pY576

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IP: FAK (C20)

FAK

pY118-paxillin

IB:

paxillin

pY31-paxillin

Mock β4GalNAc-T3C

1.0 1.8

1.0 0.5

PY20(anti-pY)

1.0 3.5

Dβ4GalNAc-T3Mock

1.0 1.8

pERK1/2

ERK1/2

β-actin

WCL

ILKIntegrin-Linked Kinase

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Integrin-Linked Kinase binding proteins

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The Journal of Cell Biology, Volume 155, Number 4, November 12, 2001 505-510

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ILK-PINCH-parvin (IPP) complex

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Signaling through IPP complex

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Activation of MAP kinasecascades by integrins

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Activation of MAP kinase cascades by integrins

Modified from Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Focal Adhesion Kinase

ERKERK

JNK

CrkII

C3G

Rap-1

B-Raf

Raf

Shc

survival

contractibility

P P

caveolin

RasRas

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Integrin regulates actin filamentsformation via Rho GTPases

(Rho, Rac, CDC42)

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Integrin signaling to the cytoskeleton via Rho GTPases

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Rho Rac CDC42

cofilinP

STRESS FIBERS

Lamellipodia

FILOPODIA

Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Mechanism of activation of N-WASP

P Active N-WASP

45Adapted from Journal of Cell Science 114, 1801-1809, 2001

Mechanism of activation of WAVE2

46Journal of Cell Science 114, 1801-1809, 2001

Cooperativity between growth-factor receptors and integrins

NATURE CELL BIOLOGY VOL 2 DECEMBER 2000 E225-229

growth factor receptorintegrin

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1

2

3

integrin Receptor Tyrosine Kinase

Integrin regulates signaling in RTK/Ras/MAPK pathway

cytoplasm

nucleus

Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

1. Activation of receptor tyrosine kinase(RTK)

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• 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

Integrin regulation of growth factor receptors

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FAK integrates growth-factor and integrinsignals to promote cell migration

51Current Opinion in Genetics & Development 2004, 14:92–101

2. Coupling between upstream and downstream molecules

Between Ras-Raf

– EGFR, PDGF-R in

anchorage-dependent

cell growth

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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 cytoskeleton-

dependent (NIH 3T3 cells)

– G1 phase progression

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Examples of synergisms between integrins and RTK signaling

Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323