title: The large conductance calcium-activated potassium channel affects extrinsic and intrinsic mechanisms of apoptosis.
abstract: The large-conductance calcium-activated K(+) or BK channel underlies electrical signals in a number of different cell types. Studies show that BK activity can also serve to regulate cellular homeostasis by protecting cells from apoptosis resulting from events such as ischemia. Recent coimmunoprecipitation studies, combined with mass spectrometry, suggest putative protein partners that interact with BK to regulate intrinsic and extrinsic apoptotic pathways. This study tests two of those partners to determine the effects on these two signaling pathways. Through reciprocal coimmunoprecipitation (coIP) experiments, we show that BK interacts with p53 and fas-associated protein with death domain (FADD) in mouse brain and when overexpressed in a heterologous expression system, such as HEK293 cells. Moreover, coIP experiments with N- and C-terminal fragments reveal that FADD interacts with the C-terminus of BK, whereas p53 interacts with either the N- or the C-terminus. Immunolocalization studies show that BK colocalizes with p53 and FADD in the mitochondrion and plasmalemma, respectively. HEK cells that stably express BK are more resistant to apoptosis when p53 or FADD is overexpressed or when their intrinsic and extrinsic pathways are stimulated via mitomycin C or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), respectively. Moreover, when stimulating with TRAIL, caspase-8 activation decreases in BK-expressing cells. These data suggest that BK is part of a larger complex of proteins that protects against apoptosis by interacting with proapoptotic proteins, such as p53 and FADD.
authors.affiliation: Department of Otolaryngology-HNS, University of South Florida Morsani College of Medicine, Tampa, Florida.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Sakai
authors.forename: Bernd
authors.initials: B
authors.lastname: Sokolowski
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology*; D001921:Brain / Q000378:metabolism; D053181:Caspase 8; D003001:Cloning, Molecular; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D051336:Mitochondrial Membranes / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D053200:Fas-Associated Death Domain Protein; D051036:Large-Conductance Calcium-Activated Potassium Channels; D011108:Polymers; D016159:Tumor Suppressor Protein p53; C082201:plasmalene; D053181:Caspase 8
keywords: BK channel; FADD; apoptosis; p53; protein-protein interactions