TY - JOUR
T1 - The Rho-guanine nucleotide exchange factor Trio controls leukocyte transendothelial migration by promoting docking structure formation
AU - Van Rijssel, Jos
AU - Kroon, Jeffrey
AU - Hoogenboezem, Mark
AU - Van Alphen, Floris P.J.
AU - De Jong, Renske J.
AU - Kostadinova, Elena
AU - Geerts, Dirk
AU - Hordijk, Peter L.
AU - Van Buul, Jaap D.
PY - 2012/8/1
Y1 - 2012/8/1
N2 - Leukocyte transendothelial migration involves the active participation of the endothelium through the formation of apical membrane protrusions that embrace adherent leukocytes, termed docking structures. Using live-cell imaging, we find that prior to transmigration, endothelial docking structures form around 80% of all neutrophils. Previously we showed that endothelial RhoG and SGEF control leukocyte transmigration. In this study, our data reveal that both full-length Trio and the first DH-PH (TrioD1) domain of Trio, which can activate Rac1 and RhoG, interact with ICAM-1 and are recruited to leukocyte adhesion sites. Moreover, upon clustering of ICAM-1, the Rho-guanine nucleotide exchange factor Trio activates Rac1, prior to activating RhoG, in a filamin-dependent manner. We further show that docking structure formation is initiated by ICAM-1 clustering into ring-like structures, which is followed by apical membrane protrusion. Interestingly, we find that Rac1 is required for ICAM-1 clustering, whereas RhoG controls membrane protrusion formation. Finally, silencing endothelial Trio expression or reducing TrioD1 activity without affecting SGEF impairs both docking structure formation and leukocyte transmigration. We conclude that Trio promotes leukocyte transendothelial migration by inducing endothelial docking structure formation in a filamin-dependent manner through the activation of Rac1 and RhoG.
AB - Leukocyte transendothelial migration involves the active participation of the endothelium through the formation of apical membrane protrusions that embrace adherent leukocytes, termed docking structures. Using live-cell imaging, we find that prior to transmigration, endothelial docking structures form around 80% of all neutrophils. Previously we showed that endothelial RhoG and SGEF control leukocyte transmigration. In this study, our data reveal that both full-length Trio and the first DH-PH (TrioD1) domain of Trio, which can activate Rac1 and RhoG, interact with ICAM-1 and are recruited to leukocyte adhesion sites. Moreover, upon clustering of ICAM-1, the Rho-guanine nucleotide exchange factor Trio activates Rac1, prior to activating RhoG, in a filamin-dependent manner. We further show that docking structure formation is initiated by ICAM-1 clustering into ring-like structures, which is followed by apical membrane protrusion. Interestingly, we find that Rac1 is required for ICAM-1 clustering, whereas RhoG controls membrane protrusion formation. Finally, silencing endothelial Trio expression or reducing TrioD1 activity without affecting SGEF impairs both docking structure formation and leukocyte transmigration. We conclude that Trio promotes leukocyte transendothelial migration by inducing endothelial docking structure formation in a filamin-dependent manner through the activation of Rac1 and RhoG.
KW - Animals
KW - COS Cells
KW - Cell Adhesion
KW - Cell Tracking
KW - Chlorocebus aethiops
KW - Contractile Proteins/chemistry
KW - Electric Impedance
KW - Filamins
KW - Guanine Nucleotide Exchange Factors/chemistry
KW - HEK293 Cells
KW - HeLa Cells
KW - Humans
KW - Intercellular Adhesion Molecule-1/metabolism
KW - Leukocytes/cytology
KW - Microfilament Proteins/chemistry
KW - Protein Serine-Threonine Kinases/chemistry
KW - Protein Structure, Tertiary
KW - Transendothelial and Transepithelial Migration
KW - rac1 GTP-Binding Protein/metabolism
KW - rho GTP-Binding Proteins/chemistry
UR - http://www.scopus.com/inward/record.url?scp=84864750941&partnerID=8YFLogxK
U2 - 10.1091/mbc.E11-11-0907
DO - 10.1091/mbc.E11-11-0907
M3 - Article
C2 - 22696684
AN - SCOPUS:84864750941
VL - 23
SP - 2831
EP - 2844
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 15
ER -