Transcription initiation by RNA polymerase II (RNAP II) requires distinct classes of transcription factors. These include (i) general transcription factors (GTFs) that are required for initiation at all or most promoters; (ii) promoter-specific factors, including both activators and repressors, that affect the rate of transcription; and (iii) cofactors that mediate either activation or repression. The GTFs include TBP (TATA binding protein), TFIIB, TFIIE, TFIIF and TFIIH, and are required for accurate initiation by RNAP II in vitro. The GTFs are also the direct targets of activators and repressors, as well as their cofactors. A remarkable feature of the GTFs is that they are highly conserved in structure and function among eukaryotic organisms. This expands the repertoire of experimental approaches available to study transcriptional mechanisms. The yeast Saccharomyces cerevisiae is an especially valuable tool for such studies because of its ability to be manipulated by extraordinarily powerful genetic methods. Accordingly, yeast genetics can be used to investigate mechanisms controlling transcription, with the outcome relevant for understanding transcriptional control in higher organisms. The objective of this proposal is to characterize the mechanisms that regulate eukaryotic gene expression, focusing on the roles played by the GTFs. A powerful combination of genetic and biochemical methods are proposed to study the processes affecting both the accuracy and rate of transcription by RNA pol II in yeast The proposal will focus on the roles of TFIIB and a novel protein, designated Ssu72, that interacts with TFIIB.
Specific Aims are (I) to define the mechanism of transcription start site selection; (2) to define the role of TFIIB in transcriptional activation; (3) to define the role of Ssu72 in both the accuracy and rate of transcription; and (4) to identify additional factors that affect either the rate or accuracy of initiation. The health relatedness of this proposal is readily apparent since gene expression is regulated primarily at the level of transcription and many human diseases, especially many forms of cancer, are known to be a consequence of aberrant gene expression.
| Lamas-Maceiras, Mónica; Singh, Badri Nath; Hampsey, Michael et al. (2016) Promoter-Terminator Gene Loops Affect Alternative 3'-End Processing in Yeast. J Biol Chem 291:8960-8 |
| Olayanju, Bola; Hampsey, James Jensen; Hampsey, Michael (2015) Genetic analysis of the Warburg effect in yeast. Adv Biol Regul 57:185-92 |
| Singh, Badri Nath; Hampsey, Michael (2014) Detection of short-range chromatin interactions by chromosome conformation capture (3C) in yeast. Methods Mol Biol 1205:209-18 |
| Rosado-Lugo, Jesús D; Hampsey, Michael (2014) The Ssu72 phosphatase mediates the RNA polymerase II initiation-elongation transition. J Biol Chem 289:33916-26 |
| Yadon, Adam N; Singh, Badri Nath; Hampsey, Michael et al. (2013) DNA looping facilitates targeting of a chromatin remodeling enzyme. Mol Cell 50:93-103 |
| Hampsey, Michael (2012) Molecular biology. A new direction for gene loops. Science 338:624-5 |
| Goel, Shivani; Krishnamurthy, Shankarling; Hampsey, Michael (2012) Mechanism of start site selection by RNA polymerase II: interplay between TFIIB and Ssl2/XPB helicase subunit of TFIIH. J Biol Chem 287:557-67 |
| Hampsey, Michael; Singh, Badri Nath; Ansari, Athar et al. (2011) Control of eukaryotic gene expression: gene loops and transcriptional memory. Adv Enzyme Regul 51:118-25 |
| Seibold, Steve A; Singh, Badri Nath; Zhang, Chunfen et al. (2010) Conformational coupling, bridge helix dynamics and active site dehydration in catalysis by RNA polymerase. Biochim Biophys Acta 1799:575-87 |
| Laine, Jean-Philippe; Singh, Badri Nath; Krishnamurthy, Shankarling et al. (2009) A physiological role for gene loops in yeast. Genes Dev 23:2604-9 |
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