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Conserved BK Channel-Protein Interactions Reveal Signals Relevant to Cell Death and Survival

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dc.contributor.author Sokolowski, Bernd
dc.contributor.author Orchard, Sandra
dc.contributor.author Harvey, Margaret
dc.contributor.author Sridhar, Settu
dc.contributor.author Sakai, Yoshihisa
dc.date.accessioned 2012-02-27T08:56:30Z
dc.date.available 2012-02-27T08:56:30Z
dc.date.issued 2011-12-09
dc.identifier.citation PLoS ONE 6(12): e28532 en
dc.identifier.issn 1932-6203
dc.identifier.uri http://dx.doi.org/10.1371/journal.pone.0028532
dc.identifier.uri http://hdl.handle.net/1956/5637
dc.description.abstract The large-conductance Ca2+-activated K+ (BK) channel and its b-subunit underlie tuning in non-mammalian sensory or hair cells, whereas in mammals its function is less clear. To gain insights into species differences and to reveal putative BK functions, we undertook a systems analysis of BK and BK-Associated Proteins (BKAPS) in the chicken cochlea and compared these results to other species. We identified 110 putative partners from cytoplasmic and membrane/cytoskeletal fractions, using a combination of coimmunoprecipitation, 2-D gel, and LC-MS/MS. Partners included 14-3-3c, valosin-containing protein (VCP), stathmin (STMN), cortactin (CTTN), and prohibitin (PHB), of which 16 partners were verified by reciprocal coimmunoprecipitation. Bioinformatics revealed binary partners, the resultant interactome, subcellular localization, and cellular processes. The interactome contained 193 proteins involved in 190 binary interactions in subcellular compartments such as the ER, mitochondria, and nucleus. Comparisons with mice showed shared hub proteins that included N-methyl-Daspartate receptor (NMDAR) and ATP-synthase. Ortholog analyses across six species revealed conserved interactions involving apoptosis, Ca2+ binding, and trafficking, in chicks, mice, and humans. Functional studies using recombinant BK and RNAi in a heterologous expression system revealed that proteins important to cell death/survival, such as annexinA5, cactin, lamin, superoxide dismutase, and VCP, caused a decrease in BK expression. This revelation led to an examination of specific kinases and their effectors relevant to cell viability. Sequence analyses of the BK C-terminus across 10 species showed putative binding sites for 14-3-3, RAC-a serine/threonine-protein kinase 1 (Akt), glycogen synthase kinase-3b (GSK3b) and phosphoinositide-dependent kinase-1 (PDK1). Knockdown of 14-3-3 and Akt caused an increase in BK expression, whereas silencing of GSK3b and PDK1 had the opposite effect. This comparative systems approach suggests conservation in BK function across different species in addition to novel functions that may include the initiation of signals relevant to cell death/survival. en
dc.language.iso eng en
dc.publisher Public Library of Science en
dc.rights Copyright 2011 Sokolowski et al. en
dc.rights.uri http://creativecommons.org/licenses/by/2.5/ en
dc.title Conserved BK Channel-Protein Interactions Reveal Signals Relevant to Cell Death and Survival en
dc.type Peer reviewed en
dc.type Journal article en
dc.subject.nsi VDP::Mathematics and natural science: 400::Basic biosciences: 470::Cell biology: 471 en
dc.type.version publishedVersion en


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