Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10802442 | Biochimica et Biophysica Acta (BBA) - Molecular Cell Research | 2013 | 9 Pages |
Abstract
A1/Bfl-1 is a NF-κB dependent, anti-apoptotic Bcl-2 family member that contains four Bcl-2 homology domains (BH) and an amphipathic C-terminal domain, and is expressed in endothelial cells (EC). Based on NF-κB reporter assays in bovine aortic EC, we have previously demonstrated that A1, like Bcl-2 and Bcl-xL, inhibits NF-κB activation. These results, however, do not fully translate when evaluating the cell's own NF-κB machinery in human EC overexpressing A1 by means of recombinant adenovirus (rAd.) mediated gene transfer. Indeed, overexpression of full-length A1 in human umbilical vein EC (HUVEC), and human dermal microvascular EC (HDMEC) failed to inhibit NF-κB activation. However, overexpression of a mutant lacking the C-terminal domain of A1 (A1ÎC) demonstrated a potent NF-κB inhibitory effect in these cells. Disparate effects of A1 and A1ÎC on NF-κB inhibition in human EC correlated with mitochondrial (A1) versus non-mitochondrial (A1ÎC) localization. In contrast, both full-length A1 and A1ÎC protected EC from staurosporine (STS)-induced cell death, indicating that mitochondrial localization was not necessary for A1's cytoprotective function in human EC. In conclusion, our data uncover a regulatory role for the C-terminal domain of A1 in human EC: anchoring A1 to the mitochondrion, which conserves but is not necessary for its cytoprotective function, or by its absence freeing A1 from the mitochondrion and uncovering an additional anti-inflammatory effect.
Keywords
HDMECHUVECEBM-2RT-PCRICAM-1Endothelial Basal Medium-2RLUSTSBAECPACHEK 293ECLhemagglutinin ARenal proximal tubular epithelial cellsTNFFACSRPTECRSVCHXMOIstaurosporinehuman umbilical vein ECIHCImmunohistochemistryanalysis of varianceANOVAApoptosisstandard error of meanC-terminal domainfluorescence-activated cell sortingEndothelial cellscycloheximideVascular endothelial growth factorVascular Endothelial Growth Factor (VEGF)tumor necrosis factornuclear factor kappa-BSEMintercellular adhesion molecule-1Mitochondriarelative light unitsreverse transcription polymerase chain reactionWestern blotRous sarcoma virusmultiplicity of infection
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Renata P. Guedes, Eduardo Rocha, Jerome Mahiou, Herwig P. Moll, Maria B. Arvelo, Janis M. Taube, Clayton R. Peterson, Elzbieta Kaczmarek, Christopher R. Longo, Cleide G. da Silva, Christiane Ferran,