McDermott, John CharlesKelebeev, Jonathan2023-03-282023-03-282022-11-102023-03-28http://hdl.handle.net/10315/40981Hippo signalling is a prominent regulator of cell proliferation, differentiation, and death. Previously, we characterized the Hippo transcriptional effector TAZ as a repressor of the myogenic differentiation program. Without DNA-binding ability, TAZ function exclusively depends on its protein:protein interaction network. This prompted us to undertake a proteomic-based study aimed at identifying the TAZ interactome in striated muscle cells. Using a novel GFP-Nanotrap based affinity purification approach coupled with LC-MS/MS protein identification, we document a comprehensive list of known and novel TAZ interactome components in myogenic cells. TAZ interacting proteins include components of Hippo signalling: TEAD1-4, LATS1, 14-3-3 proteins. Epigenetic regulators were also represented: NuRD complex, FACT complex, and SWI/SNF complex. We focused on characterizing the TAZ interaction with the Wnt co-repressor TLE3 in myogenic cells. In myogenic cells, TAZ and TLE3 interact and co-localize within the nucleus. Functionally, TAZ and TLE3 repress β-catenin activation, as indicated by the Wnt-responsive TOP FLASH reporter gene system. Depletion of TAZ reduced the degree of TLE3-mediated repression of β-catenin activation. TAZ and TLE3 repressed MyoD-driven activation of the myogenin promoter. Overall, these data demonstrate a role for a TAZ/TLE3 complex in repressing the myogenic differentiation machinery having implications for muscle development and regeneration.Author owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.Molecular biologyBiochemistryCellular biologyElectronic Thesis or Dissertation2023-03-28TAZMuscleStriatedSkeletalProteinInteractionNetworkHippoSignalling