Role of the actin cytoskeleton in G-protein-coupled receptor activation of PYK2 and paxillin in vascular smooth muscle

被引:37
作者
Ohanian, V [1 ]
Gatfield, K [1 ]
Ohanian, J [1 ]
机构
[1] Univ Manchester, Manchester Royal Infirm, Dept Med, Manchester M13 9WL, Lancs, England
关键词
kinase; mesenteric arteries; phosphorylation;
D O I
10.1161/01.HYP.0000167990.82235.3c
中图分类号
R6 [外科学];
学科分类号
1002 ; 100210 ;
摘要
Dynamic remodeling of the actin cytoskeleton occurs during agonist-induced smooth muscle contraction. Tyrosine phosphorylation of the adaptor protein paxillin has been implicated in regulation of actin filament formation and force development. We have investigated the role of the actin cytoskeleton in noradrenaline ( NA)-induced and endothelin ( ET)-induced activation of the calcium-dependent nonreceptor tyrosine kinase PYK2 and subsequent phosphorylation of paxillin in rat small mesenteric arteries. NA and ET induced a rapid and prolonged activation of PYK2, as shown by increased phosphorylation at Y402 and Y881, and a concomitant association of the kinase with a Triton X-100 insoluble membrane ( cytoskeleton) compartment. Both agonists also increased phosphorylation of paxillin at Y31 and Y118 with a similar time course as PYK2 phosphorylation, and induced its association with the same membrane compartment as PYK2. Treatment of arteries with cytochalasin D disrupted stress fibers and inhibited NA-induced and ET-induced force in a myosin light chain 20 phosphorylation independent and reversible manner. However, cytochalasin D treatment had no effect on NA-induced and ET-induced phosphorylation of either PYK2 or paxillin but did prevent their association with the TritonX-100 insoluble membrane compartment. These results show that in mesenteric arteries an intact cytoskeleton and force development are not prerequisites for G-protein-coupled receptor-induced activation of PYK2 and paxillin, by tyrosine phosphorylation, in vascular tissue, but are necessary for the translocation of PYK2 and paxillin to the membrane.
引用
收藏
页码:93 / 99
页数:7
相关论文
共 30 条
[1]   Stiffness changes in cultured airway smooth muscle cells [J].
An, SS ;
Laudadio, RE ;
Lai, J ;
Rogers, RA ;
Fredberg, JJ .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2002, 283 (03) :C792-C801
[2]   RAFTK/Pyk2-mediated cellular signalling [J].
Avraham, H ;
Park, SY ;
Schinkmann, K ;
Avraham, S .
CELLULAR SIGNALLING, 2000, 12 (03) :123-133
[3]   Exchange of the actin-bound nucleotide in intact arterial smooth muscle [J].
Bárány, M ;
Barron, JT ;
Gu, LP ;
Bárány, K .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (51) :48398-48403
[4]   CHARACTERIZATION OF TYROSINE PHOSPHORYLATION OF PAXILLIN IN-VITRO BY FOCAL ADHESION KINASE [J].
BELLIS, SL ;
MILLER, JT ;
TURNER, CE .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (29) :17437-17441
[5]   Regulation of a calcium-dependent tyrosine kinase in vascular smooth muscle cells by angiotensin II and platelet-derived growth factor - Dependence on calcium and the actin cytoskeleton [J].
Brinson, AE ;
Harding, T ;
Diliberto, PA ;
He, YQ ;
Li, X ;
Hunter, D ;
Herman, B ;
Earp, HS ;
Graves, LM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (03) :1711-1718
[6]   Rho-stimulated contractility drives the formation of stress fibers and focal adhesions [J].
ChrzanowskaWodnicka, M ;
Burridge, K .
JOURNAL OF CELL BIOLOGY, 1996, 133 (06) :1403-1415
[7]   Vascular smooth muscle actin cytoskeleton in cerebral artery forced dilatation [J].
Cipolla, MJ ;
Osol, G .
STROKE, 1998, 29 (06) :1223-1228
[8]   Signal Transduction in smooth muscle - Invited review: Focal adhesion and small heat shock proteins in the regulation of actin remodeling and contractility in smooth muscle [J].
Gerthoffer, WT ;
Gunst, SJ .
JOURNAL OF APPLIED PHYSIOLOGY, 2001, 91 (02) :963-972
[9]   SMALL ARTERY STRUCTURE IN HYPERTENSION - DUAL PROCESSES OF REMODELING AND GROWTH [J].
HEAGERTY, AM ;
AALKJAER, C ;
BUND, SJ ;
KORSGAARD, N ;
MULVANY, MJ .
HYPERTENSION, 1993, 21 (04) :391-397
[10]  
Hiregowdara D, 1997, J BIOL CHEM, V272, P10804