1.

2. Jak-STAT Janus STAT

3. - - - -

4. -

12o

Stat-3

388

:

1. , (RTK: receptors tyrosine kinases). .

2. . , (associated tyrosine kinase), . . , , , . .

, - - .

1.

(

389

. , , . , .

, .

12.2

. .

. , IL-4, IL-5, IL-6 IL-10, - -.

- -, . - -, . . , - -.

, ( MHC II -) - - -2 (IL-2). IL-2 , . , , -, -, -. - , -. , , .

390

12.2 . C- -. -. , L-2. -, .

F (tumor necrosis factor) -, -.

(Kd 10-10-10-12 M). picomoles.

, , :

1. IL-1, .

2. ( ), IL2-9, IL11-13, GM-CSF (granulocyte-macrophage colony stimulating factor), G-CSF (granulocyte colony stimulating factor), LIF (leukemia inhibitory factor), CNTF (ciliary neurotrophic factor), OSM (oncostatine), . 4 , WSXWS (W: , S: , : ).

3. ( ) , , IL-10. 4 , WSXWS.

4. TNF , CD40 Fas.

-

391

, , GPCRs.

12.3 ,

. () IL-1 . . () ( ). () ( ). () TNF , CD40 Fas.

2 3 . - C- .

- . , - IL-2 IL-4, IL-7 IL-9. gp130 -.

, , . IL-6 - gp130 . : , - - - .

392

12.4 -, Epo-R G-

CSFR, - IL-6, IL-2, IL-4 LIF (leukemia inhibitory factor). , gp130, -.

, , . :

1. . , .

2. , - , .

, , . Src Jak.

Lck (p56lck) Src, IL-2. -2 (IL-2) , , . 2 , . Lck IL-2 . - Lck , .

Lck, IL-2, Fyn Jak1 Jak3 (Janus kinases).

393

, .

Jak1Jak2

Lck

PI3-K

c-myc c-fosPLC

12.5 2. IL2 Jak1 Jak3. - Stat5. , Shc, SH2 , Ras. Stat5 . IL-2 Lck Syk, c-Myc c-Fos.

, . SH2 , . 3- Shc , Ras . Ras Shc/Grb2/mSos.

.

Jak-Stat Ras MAP C 3-

394

2. Jak-Stat

Jak-Stat . .

Janus

Janus (Jaks). Janus () , Jak , , . , JAK Just another kinase. Jak (Jak1, Jak2, Jak3, Jak4). Jak . , . C

12.6 Jak . JH1 (JH: Janus kinase homology region)

. JH2 . - Jak.

Jak

, , Box1 Box2. , Jak . Jak - .

12.7 Jak

. , Jak trans-. Jak . .

C D E JH1 JH2

395

( ) Jak . Jak 6 (Jak1 Jak2), (Jak1 Tyk1). , Jak STAT, .

12.8 IL-6, Jak1 Jak2, INF, Jak1 Tyk. , Jak STAT, , ISRE (interferon stimulated response element) - .

STAT

STAT , . 7 STAT : STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b STAT6. . , , , , , , . . STATs , 50 STATs .

IL-6

IFN

IL-6 IFN

396

STAT : STAT STATs : ,

, N (-domain), coiled coil , STAT DNA (DNA domain), (LD: linker domain) - SH2 . STATs ().

STATs (S) TAD (transcriptional activation domain) COOH .

NH2- -COOH

TAD

12.9 STATs (). T STAT (). (a) - STAT4. (- -). (b) STAT1

GAS DNA. coiled-coil , DNA binding domains, SH2 .

STATs

STAT -1. 1 (IFN IFN) - , STATs. o IFNa, IFN-stimulated gene response element (ISRE), ISGF3 (IFN-stimulated gene factor 3). o IFN-stimulated gene response element (GAS) , , GAF (gamma-activated factor). , ISGF3 GAF STATs. ISGF3 STAT1, STAT2 p48/ISGF3/IRF9 ( STAT, IFN-regulatory factors) GAF STAT1.

7 STAT : STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b STAT6. , .

ND Coiled-coil DNA LD SH2 Y S

P P

397

STATs STATs , SH2 ,

JAK. . STATs . STATs , DNA ( interferon stimulated response element gamma activation site), . STAT2, STATs .

12.10 STATs, JAK . orvath C.M., Stat proteins and transcriptional responses to extracellular signals, Trends in Biochem. Sci., 2000, 25, 496-502

STATs STTs GTP Ran (importin), , . STAT NLS (nuclear localization sequence), . GTP Ran, , . , Ran GTP.

STAT DNA

- - STAT GAS (gamma activation sites) DNA, --. GAS (gene promoters), PIE: prolactin-inducible element .

- STAT1-STA2 p48/ISGF3, ISRE DNA. STAT - GAS.

STATs

STAT

398

STATs DNA, enhancer binding protein ( SP-1, GR), STAT .

DNA STATs TAD, -, .

12.11 STATs, DNA. () STAT

GAS (gamma activation sites) DNA, --. () - STAT1-STA2 p48/ISGF3, ISRE DNA. () STAT - GAS. () STATs DNA, enhancer binding protein ( SP-1, GR), STAT . () DNA STATs TAD, -, .

orvath C.M., Stat proteins and transcriptional responses to extracellular signals, Trends in Biochem. Sci., 2000, 25, 496-502

() - ()

()

/

() DNA

() GAS element

/ RNA

399

STAT STAT :

1. NES 2. , STAT

DNA . DNA, STAT1 NES, , Ran.

STATs

Stat1 Stat. Stat1 , , IFNs. Stat2 Stat GAS DNA. Stat2 , , . IFN/ Stat1. Stat3 , , . knock-out . Stat3 , . Stat4 IL-12. Stat4 . Stat4 B IgE. Stat5, 5 5 . Stat5 . Stat5 IL-3, IL-12 GM-CSF. . Stat5 , T . Stat6 .

IL-4 . - B . Stat6 C/EBP IgE.

3. - -

- - , . , .

400

- , , - - . - ,

- , - - . - ( ) . MHC (major histocompatibility complex) - - ( 12.2). MHC . - - . , . - -,

- , -.

- - , , .

CD3

12.12 -. Ti, CD3. ITAM. R. Goldsby, T. Kindt and J. Kuby, Immunology, Freeman editions, 5th edition, 2003.

Ti , . ,

i CD3

401

CD3, , , , . Tyr Leu (D/E)XXYXXL(X)6-8YXXL (ARAM: antigen recognition activation motif) (ITAM: immunoreceptor tyrosine-based activation motif). ARAM CD3, CD3, CD3 . ARAM -. -, Tyr ITAMs (ARAMs) , SH2 . .

- -. -. CD4 CD8, - - , . - , Src. , .

TCR (T cell Receptor) MHC- 7 ( -) CD28 ( -).

12.13 - TCR (T cell receptor).

- (THelper) - TCR CD4, - (Tcytotoxic), - CD8. - MHC-II (o CD4) MHC- (o CD8) TCR- -MHC. - -, CD28-B7, CD45R-CD22, CD2-LFA-3. R. Goldsby, T. Kindt and J. Kuby, Immunology, Freeman editions, 5th edition, 2003.

402

12.14 () - CD4 CD8. () CD4

CII- (pMHCII) TCR.

- -

- - . TCR Src ( Lck: Leucocyte specific kinase Fyn: Fgr/yes novel kinase) , . , Lck Fyn -, , CD4/8 -.

SH2 SH2 Kinase

Domain organization of ZAP-70

12.15 TCR

Lck, - CD4/8 Tyr -. The structure, regulation, and function of ZAP-70. Au-Yeung BB, Deindl S, Hsu LY, Palacios EH, Levin SE, Kuriyan J, Weiss A. The structure, regulation, and function of ZAP-70. Immunol Rev. 2009, 228(1):41-57.

. .

403

-70 ( associated protein 70: 70 kDa ), SH2 . , 70 Lck . ZAP-70 , -: L (Linker for activation of T-cells), .

LAT, Tyr, , SH2 , T-cell signalosome. Tyr132 LAT PLC, Tyr171 PI3-K , Tyr191 Grb2 , GDS (Grb-like adaptor downstream of Shc), Tyr226 Vav.

LAT adaptor protein

12.16 LAT ,

ZAP-70 (, , - ) . Houtman JC, Barda-Saad M, Samelson LE. Examining multiprotein signaling complexes from all angles, FEBS J. (2005) 272:5426-35.

SH3 ,

, T-cell signalosome. SH3 GADS SLP-76 (SH2-domain-containing leukocyte phosphoprotein of 76 kDa), Vav Itk (IL2-inducible T-cell kinase) TCR.

Itk : Itk , 3,4,5 PtdIns(3,4,5)P3. PtdIns(3,4,5)P3 PtdIns(4,5)P2 3-, Lck. TCR, Itk PtdIns(3,4,5)P3 S2 LAT. , Lck

404

SLP-76/Vav/Itk. Itk PLC- , .

12.17 TCR

Lck. Lck 3-, PtdIns(4,5)P2 PtdIns(3,4,5)P3. Itk PtdIns(3,4,5)P3 . Lck. Finkelstein LD, Schwartzberg PL. Tec kinases: shaping T-cell activation through actin, Trends Cell Biol. (2004) 14:443-51.

12.18 TCR. PLC PtdIns(4,5)P2 (2)

(IP3) (DAG). I3 - , Ca2+ . DAG Ca2+ PKC. , DAG RasGEF, RasGRP, PKC.

PC PKC, -, Ca2+ DAG.

405

NFAT (nuclear factor for activated T-cells) JNK. PKC NFB. NFAT NFB IL-2. NF - .

Vav1 . Vav1, - ( Vav2, Vav3 ). DH , Rho GTP, (Ac) Tyr, , ( Vav GEF), SH3 SH2 . , -. TCR, Vav1 Tyr174 , SH2 o -cell signalosome ( SLP-76 LAT). , Vav1 GTP Rac Cdc42, WASP (Wiskott-Aldrish syndrome protein) Arp2/3 ( . 10, .328-329). WASP signalosome Nck (non-catalytic region of tyrosine kinase), SH2 SH3 . SH2 SLP76 SH3 WASP. WASP GTP Cdc42, .

Vav1

12.19 Vav1, -

. Vav Tyr (Ac) , GTP Cdc42 DH . Tybulewicz VL., Vav-family proteins in T-cell signaling, Curr Opin Immunol. (2005) 17(3):267-74.

406

4.

. , . , , . . , (integrate) - - (Hynes, 1987).

. , , , . ( ) . - . . , , , .

12.18 , ,

().

: ,

407

,

, ( ) 20-70 . 16 - 8 . , . , . . .

(clustering) , , ( ) ( ) . .

12.19 ()

. . Quin J, Vinogradova O and Plow E, Integrin bidirectional signaling: a molecular view, PLoS Biology, 2004, 2, 726-729.

: - : ,

( ) .

- : , . , . - - , .

408

-

. , (focal adhesion sites). ( 50 ) , . . . , , , Ca2+- .

, - FAK, .

FAK (focal adhesion kinase)

, , (Tyr397). Y397 FAK SH2 , :

3- ( p85 ), -3- , . : Ser/Thr, ( ) ( Bad, -9..). , (, ) , .

Src, FAK Tyr (Tyr925), Shc, Grb2 Ras/RK, .

409

12.20 () FAK (focal adhesion kinase),

1992 Src. () FAK, (FERM homology domain), FAT , , SH2 . Schwartz M.A., Integrin signalling revisited, Trends in Cell Biology, 2001, 11, 466-470. () FAK. FAK , , - . Schlaepfer D.D., Mitra S.K., Multiple connections link FAK to cell motility and invasion, Current Opinion in Genetics and Development, 2004, 14, 92-101.

. .

.

410

12.21 (15) ()

FAK/PI3-K/PKB/BAD-Caspase9 () FAK/Src/Shc/Sos/Ras/MAPK . Gomperts et al, Signal Transduction, 2002, ELSEVIER.

12.22 FAK.

FAK . FAK Src, (Tyr925). Schlaepfer DD, Mitra SK, Ilic D. Control of motile and invasive cell phenotypes by focal adhesion kinase. Biochim Biophys Acta. 2004, 1692(2-3), 77-102.

411

FAK , / . , / .

Rho GTP. FAK p190RhoGEF, Rho. Rho ROCK, MLC (myosin light chain), MLCK ( ). GTP Rho: Tyr925 FAK Grb2, Ras/MEK/ERK Rho.

GTP Rho . Graf-GAP SH3 FAK Rho. , FAK/Src p190RhoGAP, Rho. . FAK , .

12.23 FAK

Rho. Schlaepfer D.D., Mitra S.K., Multiple connections link FAK to cell motility and invasion, Current Opinion in Genetics and Development, 2004, 14, 92-101.

412

.

. FAK .

v-Src FAK . v-Src-FAK (invadopodia), (), . Tyr397 FAK Src . , FAK-Src JNK JNK, JSAP1, p130CAS/CRK/DOCK/Rac.

JNK Rac, Sr178 2, 9. .

12.24 / FAK/ JNK

. Schlaepfer D.D., Mitra S.K., Cur. Opinion Genet Devel, 2004, 14, 92-101.

413

Darnall JE, Kerr IM and Stark GR, Jak-Stat patways and transcriptional activation in response

to IFNs and other extracellular signalling pathways, Science, 1994, 264, 1415-1420. DeMali K., Wenerberg K., Burridge K., Integrin signaling to the actin cytoskeleton, Current

Opinion in Cell Biology, 2003, 15, 572-582 Fagerholm S., Hilden T., Gahmberg C., P marks the spot: site-specific integrin

phosphorylation regulates molecular interactions, Trends in Biochemical Sciences, 2004, 29, 504-512

orvath C.M., Stat proteins and transcriptional responses to extracellular signals, Trends in Bioch. Sci., 2000, 25, 496-502

Hynes RO, Integrins: A family of cell surface receptors, Cell, 1987, 48, 549-550 Krauss G., Biochemistry of Signal Transduction and Regulation, Wiley-VCH editions, 2001 Parsons J.T., Focal adhesion kinase: the first ten years, J. Cell Science, 2003, 116, 1409-1416 Quin J, Vinogradova O and Plow E, Integrin bidirectional signaling: a molecular view, PLoS

Biology, 2004, 2, 726-729 Schwartz M.A., Integrin signalling revisited, Trends in Cell Biology, 2001, 11, 466-470 Schlaepfer D.D., Mitra S.K., Multiple connections link FAK to cell motility and invasion,

Current Opinion in Genetics and Development, 2004, 14, 92-101 Schlaepfer DD, Mitra SK, Ilic D. Control of motile and invasive cell phenotypes by focal

adhesion kinase. Biochim Biophys Acta. 2004, 1692(2-3), 77-102. Wozniiak M., Modzelewska K., Kwong L., Keely P., Focal adhesion regulation of cell behaviour,

Biochim. Bioph. Acta, 2004, 1692, 103-119

414