title: The Stability and Expression Level of Bok Are Governed by Binding to Inositol 1,4,5-Trisphosphate Receptors.
abstract: Bok is a member of the Bcl-2 protein family that governs the intrinsic apoptosis pathway, although the role that Bok plays in this pathway is unclear. We have shown previously in cultured cell lines that Bok interacts strongly with inositol 1,4,5-trisphosphate receptors (IP3Rs), suggesting that it may contribute to the structural integrity or stability of IP3R tetramers. Here we report that Bok is similarly IP3R-assocated in mouse tissues, that essentially all cellular Bok is IP3R bound, that it is the helical nature of the Bok BH4 domain, rather than specific amino acids, that mediates binding to IP3Rs, that Bok is dramatically stabilized by binding to IP3Rs, that unbound Bok is ubiquitinated and degraded by the proteasome, and that binding to IP3Rs limits the pro-apoptotic effect of overexpressed Bok. Agents that stimulate IP3R activity, apoptosis, phosphorylation, and endoplasmic reticulum stress did not trigger the dissociation of mature Bok from IP3Rs or Bok degradation, indicating that the role of proteasome-mediated Bok degradation is to destroy newly synthesized Bok that is not IP3R associated. The existence of this unexpected proteolytic mechanism that is geared toward restricting Bok to that which is bound to IP3Rs, implies that unbound Bok is deleterious to cell viability and helps explain the current uncertainty regarding the cellular role of Bok.
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210.
authors.forename: Jacqualyn J
authors.initials: JJ
authors.lastname: Schulman
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210.
authors.forename: Forrest A
authors.initials: FA
authors.lastname: Wright
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Han
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Zluhan
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210.
authors.forename: Laura M
authors.initials: LM
authors.lastname: Szczesniak
authors.affiliation: From the Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210 wojcikir@upstate.edu.
authors.forename: Richard J H
authors.initials: RJ
authors.lastname: Wojcikiewicz
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D055550:Protein Stability; D059748:Proteolysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: C503049:BOK protein, human; D053496:Inositol 1,4,5-Trisphosphate Receptors; C025389:Itpr1 protein, mouse; D019253:Proto-Oncogene Proteins c-bcl-2; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium
keywords: B-cell lymphoma 2 (Bcl-2) family; apoptosis; inositol trisphosphate receptor (InsP3R); proteasome; ubiquitin