Coordinated regulation and colocalization of alphav integrin and its activating enzyme proprotein convertase PC5 in vivo.
Integrin alphav is involved in intracellular-extracellular signaling important for cytoskeleton alterations and control of cell movement. In vitro experiments indicate that the integrin alphav-subunit undergoes post-translational endoproteolytic cleavage. This type of activation requires the presence of suitable kexin/subtilisin-like proprotein convertases. In vitro experiments have demonstrated that, among several proprotein convertases, PC5A, and to a threefold lesser extent furin, can activate alphav integrin. The biological significance of these in vitro data would be further supported by a coexpression and coordinated regulation of the gene expression of alphav integrin and its activating enzyme PC5 in vivo. In the present study we investigated the regulation of alphav integrin and PC5 following balloon injury in vivo. Comparative immunocytochemistry revealed a coordinated regulation of alphav integrin and PC5 during vascular remodeling in rodents. Integrin alphav was found to be upregulated in PCNA-positive, proliferating vascular smooth muscle cells. Northern blots revealed no significant regulation of furin mRNA, whereas PC5A mRNA increased during vascular remodeling, suggesting that PC5 is the major convertase during neointima formation in vivo. Incubation of vascular smooth muscle cells with the Golgi-disturbing agent brefeldin A inhibited alphav integrin maturation, indicating that endoproteolytic cleavage occurs in the trans-Golgi network, were PC5 is localized. Thus, the present study further supports the concept that activation of alphav integrin occurs in the trans-Golgi network in vascular smooth muscle cells and involves PC5.
Stawowy
Philipp
P
Department of Medicine/Cardiology, Deutsches Herzzentrum Berlin, Augustenburger Platz 1, 13353, Berlin, Germany. stawowy@dhzb.de
Graf
Kristof
K
Goetze
Stephan
S
Roser
Mattias
M
Chrétien
Michel
M
Seidah
Nabil G
NG
Fleck
Eckart
E
Marcinkiewicz
Mieczyslaw
M
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000293:injuries; D045683:Furin / Q000378:metabolism; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D039564:Integrin alphaV / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000293:injuries; D018919:Neovascularization, Physiologic; D045702:Proprotein Convertase 5 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution; D021601:trans-Golgi Network
D039564:Integrin alphaV; D012333:RNA, Messenger; D045702:Proprotein Convertase 5; D045683:Furin