Programmed cell death, or apoptosis, is a cellular process essential for normal development and tissue homeostasis. Disruption of the delicate balance between cellular life and death can lead to a host of human diseases, including cancer. BCL-2 family proteins are key apoptotic regulators and are composed of anti- and pro-apoptotic members. Pro-apoptotic BAX is a critical gatekeeper of mitochondrial apoptosis. Upon activation by cellular stress, BAX transforms from an inactive cytosolic monomer into a toxic pore that disrupts the mitochondrial membrane and initiates the apoptotic pathway. The Walensky laboratory previously identified a trigger site or """"""""on switch"""""""" at BAX's N-terminal surface. Engagement of this interaction site by specific members of the BH3-only subgroup of BCL-2 proteins initiates BAX activation, including its translocation from the cytosol to the mitochondria membrane. This year, we determined that the C-terminal groove of BAX is also compatible with BH3-only protein interaction. We believe this newly identified C-terminal trigger site propels the activation and oligomerization of BAX at the mitochondrial membrane. Here, I propose to develop novel probes to dissect the binding determinants of the two BH3-binding sites on BAX and determine how their selective triggering functionally impacts the BAX activation pathway. Specifically, I aim to (1) : Define the binding determinants for selective engagement of the N- and C-terminal trigger sites of BAX using Stabilized Alpha-Helices of BCL-2 domains (SAHBs) and (2) Dissect the functional roles of the two BH3-binding sites on BAX in triggering its direct activation. By applying multidisciplinary approaches that span chemistry, apoptosis biology, and developmental cancer therapeutics, I hope to advance our understanding of the biochemical mechanism that drives BAX-mediated apoptosis and inform the development of a new strategy to reactivate apoptosis in cancer through direct BAX activation.
BAX is a pro-apoptotic BCL-2 family member that, in response to cellular stress, transforms from an inactive cytosolic monomer into a toxic mitochondrial pore, a mechanism effectively suppressed by cancer cells to ensure their immortality. The Walensky laboratory recently identified two trigger sites at BAX's N- and C-terminal surfaces that are believed to initiate and propel the BAX activation mechanism. Using multidisciplinary approaches, I am to dissect the binding determinants of these distinct regulatory sites and determine how specific ligand interaction impacts the BAX activation process, with the ultimate goal of advancing a new therapeutic strategy for reactivating the death pathway in cancer through direct BAX interaction.
| Pritz, Jonathan R; Wachter, Franziska; Lee, Susan et al. (2017) Allosteric sensitization of proapoptotic BAX. Nat Chem Biol 13:961-967 |
