GEP100/BRAG2:: Activator of ADP-ribosylation factor 6 for regulation of cell adhesion and actin cytoskeleton via E-cadherin and α-catenin

被引:59
作者
Hiroi, Toyoko [1 ]
Someya, Akimasa [1 ]
Thompson, Walter [1 ]
Moss, Joel [1 ]
Vaughan, Martha [1 ]
机构
[1] NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA
关键词
adherens junction; F-actin;
D O I
10.1073/pnas.0604091103
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
GEP(100) (p100) was identified as an ADP-ribosylation factor (ARF) guanine nucleotide-exchange protein (GEP) that partially colocalized with ARF6 in the cell periphery. p100 preferentially accelerated guanosine 5[gamma-thio]triphosphate (GTP gamma S) binding by ARF6, which participates in protein trafficking near the plasma membrane, including receptor recycling, cell adhesion, and cell migration. Here we report that yeast two-hybrid screening of a human fetal brain cDNA library using p100 as bait revealed specific interaction with alpha-catenin, which is known as a regulator of adherens junctions and actin cytoskeleton remodeling. Interaction of p100 with alpha-catenin was confirmed by coimmunoprecipitation of the endogenous proteins from human HepG2 or CaSki cells, although colocalization was difficult to demonstrate microscopically. alpha-Catenin enhanced GTP gamma S binding by ARF6 in vitro in the presence of p100. Depletion of p100 by small interfering RNA (siRNA) treatment in HepG2 cells resulted in E-cadherin content 3-fold that in control cells and blocked hepatocyte growth factor-induced redistribution of E-cadherin, consistent with a known role of ARF6 in this process. F-actin was markedly decreased in normal rat kidney (NRK) cells overexpressing wild-type p100, but not its GEP-inactive mutants, also consistent with the conclusion that p100 has an important role in the activation of ARF6 for its functions in both E-cadherin recycling and actin remodeling.
引用
收藏
页码:10672 / 10677
页数:6
相关论文
共 49 条
[1]   ARF6 regulates a plasma membrane pool of phosphatidylinositol(4,5)bisphosphate required for regulated exocytosis [J].
Aikawa, Y ;
Martin, TFJ .
JOURNAL OF CELL BIOLOGY, 2003, 162 (04) :647-659
[2]   ADP-ribosylation factor 6 and endocytosis at the apical surface of Madin-Darby canine kidney cells [J].
Altschuler, Y ;
Liu, SH ;
Katz, L ;
Tang, K ;
Hardy, S ;
Brodsky, F ;
Apodaca, G ;
Mostov, K .
JOURNAL OF CELL BIOLOGY, 1999, 147 (01) :7-12
[3]   ADP-ribosylation factor 6 regulates actin cytoskeleton remodeling in coordination with Rac1 and RhoA [J].
Boshans, RL ;
Szanto, S ;
van Aelst, L ;
D'Souza-Schorey, C .
MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (10) :3685-3694
[4]   Phosphatidylinositol 4,5-bisphosphate and Arf6-regulated membrane traffic [J].
Brown, FD ;
Rozelle, AL ;
Yin, HL ;
Balla, T ;
Donaldson, JG .
JOURNAL OF CELL BIOLOGY, 2001, 154 (05) :1007-1017
[5]   ADP-RIBOSYLATION FACTOR, A SMALL GTP-DEPENDENT REGULATORY PROTEIN, STIMULATES PHOSPHOLIPASE-D ACTIVITY [J].
BROWN, HA ;
GUTOWSKI, S ;
MOOMAW, CR ;
SLAUGHTER, C ;
STERNWEIS, PC .
CELL, 1993, 75 (06) :1137-1144
[6]   A human exchange factor for ARF contains Sec7- and pleckstrin-homology domains [J].
Chardin, P ;
Paris, S ;
Antonny, B ;
Robineau, S ;
BeraudDufour, S ;
Jackson, CL ;
Chabre, M .
NATURE, 1996, 384 (6608) :481-484
[7]   Control of myoblast fusion by a guanine nucleotide exchange factor, loner, and its effector ARF6 [J].
Chen, EH ;
Pryce, BA ;
Tzeng, JA ;
Gonzalez, GA ;
Olson, EN .
CELL, 2003, 114 (06) :751-762
[8]   β-arrestin-mediated ADP-ribosylation factor 6 activation and β2-adrenergic receptor endocytosis [J].
Claing, A ;
Chen, W ;
Miller, WE ;
Vitale, N ;
Moss, J ;
Premont, RT ;
Lefkowitz, RJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (45) :42509-42513
[9]   Phylogenetic analysis of Sec7-domain-containing Arf nucleotide exchangers [J].
Cox, R ;
Mason-Gamer, RJ ;
Jackson, CL ;
Segev, N .
MOLECULAR BIOLOGY OF THE CELL, 2004, 15 (04) :1487-1505
[10]   Dynamics of phosphoinositides in membrane retrieval and insertion [J].
Czech, MP .
ANNUAL REVIEW OF PHYSIOLOGY, 2003, 65 :791-815