The long-term objectives of the studies proposed here are to identify and characterize factors that play an important role in transcription initiation by RNA polymerase II in vivo. We will 1) investigate the role of TFIIA in the response to oxidative stress; 2) determine the function(s) of TFIID in transcriptional processes in vivo; and 3) further characterize the transcriptional requirement for the newly defined protein SPN1. To accomplish these aims, a combination of genetic, molecular and biochemical approaches will be employed in Saccharomyces cerevisiae. Specifically, we have isolated mutant alleles of TFIIA that are defective for response to the transcriptional activator YAP1, YAP1 is the key regulator of oxidative stress in yeast, and the response to oxidative stress is conserved from yeast to humans, In fact, oxidative stress appears to play a role in a variety of human conditions, including neurodegenerative diseases, cancer, and aging. Studies described in Specific Aim 1 involve a complete biochemical and genetic characterization of the TFIIA-YAP1 connection. These results will certainly advance our understanding of the mechanism of induction of the genes required for protection from oxidative damage. In studies described in Specific Aim 2, we will map the protein-protein interactions in the TFIID complex, and exploit this information to design mutants that specifically disrupt certain TAF-TAF interactions. We will also refine our knowledge about the TFIIA-TAF40 interaction, by isolating compensatory mutants. TAF mutants generated will be characterized at the genetic and biochemical level, with the goal of determining the essential nature of the protein-protein interactions specific for TFIID functions. Using a genetic selection for suppression of an activation defective allele of TBP, we have also identified a new gene involved in transcriptional processes: SPN1. To determine the mechanisms by which this essential and highly conserved protein functions in gene expression, studies in Specific Aim 3 are focused on further characterizing SPN1, and identifying proteins that interact with SPN1 via biochemical and genetic approaches. Results generated from this proposal should provide important information on the factors and mechanisms essential for transcriptional regulation directly relevant to the human condition, since the processes of transcriptional control are conserved from yeast to humans. ? ?
| Lee, Sarah K; Fletcher, Aaron G L; Zhang, Lei et al. (2010) Activation of a poised RNAPII-dependent promoter requires both SAGA and mediator. Genetics 184:659-72 |
| Park, Young-Jun; Sudhoff, Keely B; Andrews, Andrew J et al. (2008) Histone chaperone specificity in Rtt109 activation. Nat Struct Mol Biol 15:957-64 |
| Park, Young-Jun; Sudhoff, Keely B; Andrews, Andrew J et al. (2008) Histone chaperone specificity in Rtt109 activation. Nat Struct Mol Biol : |
| Zhang, Lei; Fletcher, Aaron G L; Cheung, Vanessa et al. (2008) Spn1 regulates the recruitment of Spt6 and the Swi/Snf complex during transcriptional activation by RNA polymerase II. Mol Cell Biol 28:1393-403 |
| Stewart, Jennifer J; Fischbeck, Julie A; Chen, Xu et al. (2006) Non-optimal TATA elements exhibit diverse mechanistic consequences. J Biol Chem 281:22665-73 |
| Kraemer, Susan M; Goldstrohm, David A; Berger, Ann et al. (2006) TFIIA plays a role in the response to oxidative stress. Eukaryot Cell 5:1081-90 |
| Robinson, M M; Yatherajam, G; Ranallo, R T et al. (2005) Mapping and functional characterization of the TAF11 interaction with TFIIA. Mol Cell Biol 25:945-57 |
| Yatherajam, Gayatri; Zhang, Lei; Kraemer, Susan M et al. (2003) Protein-protein interaction map for yeast TFIID. Nucleic Acids Res 31:1252-60 |
| Fischbeck, Julie A; Kraemer, Susan M; Stargell, Laurie A (2002) SPN1, a conserved gene identified by suppression of a postrecruitment-defective yeast TATA-binding protein mutant. Genetics 162:1605-16 |
| Stargell, L A; Ogg, R C; Adkins, J N et al. (2001) Transcriptional activity of the TFIIA four-helix bundle in vivo. Proteins 43:227-32 |
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