Transcriptional activation of endoglin and transforming growth factor-beta signaling components by cooperative interaction between Sp1 and KLF6: their potential role in the response to vascular injury.
Endoglin is an endothelial membrane glycoprotein involved in cardiovascular morphogenesis and vascular remodeling. It associates with transforming growth factor-beta (TGF-beta) signaling receptors to bind TGF-beta family members, forming a functional receptor complex. Arterial injury leads to up-regulation of endoglin, but the underlying regulatory events are unknown. The transcription factor KLF6, an immediate-early response gene induced in endothelial cells during vascular injury, transactivates TGF-beta, TGF-beta signaling receptors, and TGF-beta-stimulated genes. KLF6 and, subsequently, endoglin were colocalized to vascular endothelium (ie, expressed in the same cell type) following carotid balloon injury in rats. After endothelial denudation, KLF6 was induced and translocated to the nucleus; this was followed 6 hours later by increased endoglin expression. Transient overexpression of KLF6, but not Egr-1, stimulated endogenous endoglin mRNA and transactivated the endoglin promoter. This transactivation was dependent on a GC-rich tract required for basal activity of the endoglin promoter driven by the related GC box binding protein, Sp1. In cells lacking Sp1 and KLF6, transfected KLF6 and Sp1 cooperatively transactivated the endoglin promoter and those of collagen alpha1(I), urokinase-type plasminogen activator, TGF-beta1, and TGF-beta receptor type 1. Direct physical interaction between Sp1 and KLF6 was documented by coimmunoprecipitation, pull-down experiments, and the GAL4 one-hybrid system, mapping the KLF6 interaction to the C-terminal domain of Sp1. These data provide evidence that injury-induced KLF6 and preexisting Sp1 may cooperate in regulating the expression of endoglin and related members of the TGF-beta signaling complex in vascular repair.
Botella
Luisa M
LM
Centro de Investigaciones Biológicas, Consejo Superior Investigaciones Cientificas (CSIC), Madrid, Spain. cibluisa@cib.csic.es
Sánchez-Elsner
Tilman
T
Sanz-Rodriguez
Francisco
F
Kojima
Soichi
S
Shimada
Jun
J
Guerrero-Esteo
Mercedes
M
Cooreman
Michael P
MP
Ratziu
Vlad
V
Langa
Carmen
C
Vary
Calvin P H
CP
Ramirez
Jose R
JR
Friedman
Scott
S
Bernabéu
Carmelo
C
D000818:Animals; D015703:Antigens, CD; D002339:Carotid Arteries; D002404:Catheterization / Q000009:adverse effects; D004195:Disease Models, Animal; D004357:Drug Synergism; D000071063:Endoglin; D004730:Endothelium, Vascular / Q000187:drug effects / Q000293:injuries / Q000378:metabolism; D006801:Humans; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011485:Protein Binding; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface; D015398:Signal Transduction; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D015533:Transcriptional Activation; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014471:Umbilical Veins; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*
D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; C105465:KLF6 protein, human; C491357:Klf6 protein, rat; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D016329:Sp1 Transcription Factor; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D019010:Vascular Cell Adhesion Molecule-1