The Transforming Growth Factor beta (TGFb) superfamily of ligands are highly versatile, functioning throughout development and in adult organisms as diverse as worms and humans to regulate a myriad of biological activities. TGFb family members signal through their cognate serine/threonine kinase receptors to mediate the phosphorylation and activation of receptor-regulated Smads (R-Smads), that then complex with the common Smad (co-Smad/Smad4) to transduce TGFb signals from the membrane into the nucleus. This thesis recounts the first identification of a mammalian Smad-interacting transcription factor, Foxh1. Investigation of the Smad/Foxh1 DNA-binding complex, which mediates TGFb-dependent regulation of transcription from a Gsc enhancer, determined that both Smad and Foxh1 binding sites are required. These studies also defined the first known biological difference between the highly related R-Smads, Smad2 and Smad3. Specifically, it was shown that while both can similarly participate in Smad/Foxh1 DNA-binding complexes, Smad2 activates and Smad3 represses Foxh1-mediated TGFb-dependent transcription. A detailed analysis of the Gsc enhancer element subsequently defined the sequence req irements for a functional Smad/Foxh1 enhancer (SFE). This information was utilized to direct in silico, genome wide searches for genes harbouring evolutionarily conserved SFEs, which successfully expanded the repertoire of Smad/Foxh1 targets. Analysis of these targets revealed novel roles for Smad/Foxh1 signalling in forebrain development and retinoic acid production. Finally, the importance of Foxh1 to heart development was examined. The interaction between Foxh1 and the heart specific factor Nkx2-5 was characterized with respect to TGFb-dependent regulation of Mef2c expression via a compound Foxh/Nkx2-5 enhancer (FNE). Genome-wide searches for similar FNEs identified many potential Foxh1/Nkx2-5 targets, further analysis of which will provide greater insights into how Foxh1 functions in heart development. In summary, the work presented herein expands our understanding of the role of TGFb in development through the identification and characterization of Foxh1 and its genomic targets downstream of TGFb signalling. / PhD
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/17832 |
| Date | 28 September 2009 |
| Creators | Silvestri, Cristoforo |
| Contributors | Attisano, Liliana, Medical Science |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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