Identification of a functional NF-kappa B site in the platelet endothelial cell adhesion molecule-1 promoter.
Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a type I transmembrane adhesion protein of 130 kDa that belongs to a subgroup of the Ig gene superfamily, characterized by the presence of immunoreceptor tyrosine-based inhibitory motifs. PECAM-1 is expressed in circulating platelets, monocytes, neutrophils, a selective subgroup of T cells, and in endothelial cells, where it is preferentially located at intercellular junctions and participates in leukocyte transmigratory processes. The identification of two consensus NF-kappa B sites within the PECAM-1 promoter led us to analyze their possible involvement in the PECAM-1 expression regulated by inflammatory stimuli. We found that surface expression and promoter activity of PECAM-1 in myeloid cells are regulated by modulators of NF-kappa B, including TNF-alpha, PMA, and pyrrolidine dithiocarbamate. Mobility shifts assays identified a specific NF-kappa B-binding element at +110/+120, whose mutation abolished the basal promoter activity of PECAM-1 and decreased NF-kappa B-dependent responses of the PECAM-1 gene promoter. Furthermore, cotransfection experiments with an expression vector encoding the p65 subunit of NF-kappa B showed transactivation of the PECAM-1 promoter. These results demonstrate that NF-kappa B can regulate the transcriptional activity of PECAM-1.
Botella
L M
LM
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Puig-Kröger
A
A
Almendro
N
N
Sánchez-Elsner
T
T
Muñoz
E
E
Corbí
A
A
Bernabéu
C
C
D020121:5' Untranslated Regions / Q000235:genetics; D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism* / Q000502:physiology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051996:Transcription Factor RelA; D014158:Transcription, Genetic / Q000235:genetics; D020298:U937 Cells
D020121:5' Untranslated Regions; D016328:NF-kappa B; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D051996:Transcription Factor RelA