The long term goals of this proposal are to understand the functions of the transcription factor TFIID and to identify and analyze factors with which it interacts. TFIID binds to the TATA regions of eukaryotic promoters and is required for transcription initiation by RNA polymerase II in vivo and in vitro. Mutations in the yeast SPT15 gene, which encodes TFIID, were isolated as suppressors of insertion mutations and cause a variety of mutant phenotypes and transcriptional defects. From a set of genetically characterized spt15 mutants, the sptl5 mutations will be sequenced and the mutant TFIID products will be studied biochemically. In addition, other classes of sptl5 mutations will be made in vitro and isolated in vivo. Analysis of different classes of spt15 mutants will correlate in vivo phenotypes, in vitro defects, and amino acid changes in TFIID. Analysis of spt15 mutants will also address the roles of TFIID in regulation of gene expression. To identify other components important for transcription initiation in vivo, unlinked suppressors of sptl5 mutations will be studied. One suppressor already isolated indicates that the SPT3 gene product physically interacts with TFIID. This possibility will be tested by biochemical and genetic experiments. In addition, other suppressors of sptl5 mutations will be isolated. The genes identified by these screens will be characterized to begin to elucidate their interactions with TFIID and their roles in transcription initiation. Given the conservation of transcription initiation in eukaryotes, an understanding of these functions will be relevant to understanding this process in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM045720-04
Application #
2183330
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1991-05-01
Project End
1995-04-30
Budget Start
1994-05-01
Budget End
1995-04-30
Support Year
4
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Harvard University
Department
Genetics
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Reavey, Caitlin T; Hickman, Mark J; Dobi, Krista C et al. (2015) Analysis of Polygenic Mutants Suggests a Role for Mediator in Regulating Transcriptional Activation Distance in Saccharomyces cerevisiae. Genetics 201:599-612
Neumüller, Ralph A; Gross, Thomas; Samsonova, Anastasia A et al. (2013) Conserved regulators of nucleolar size revealed by global phenotypic analyses. Sci Signal 6:ra70
Ahn, Sejin; Spatt, Dan; Winston, Fred (2012) The Schizosaccharomyces pombe inv1(+) regulatory region is unusually large and contains redundant cis-acting elements that function in a SAGA- and Swi/Snf-dependent fashion. Eukaryot Cell 11:1067-74
Rando, Oliver J; Winston, Fred (2012) Chromatin and transcription in yeast. Genetics 190:351-87
Helmlinger, Dominique; Marguerat, Samuel; Villen, Judit et al. (2011) Tra1 has specific regulatory roles, rather than global functions, within the SAGA co-activator complex. EMBO J 30:2843-52
Hickman, Mark J; Spatt, Dan; Winston, Fred (2011) The Hog1 mitogen-activated protein kinase mediates a hypoxic response in Saccharomyces cerevisiae. Genetics 188:325-38
Winston, Fred (2009) A transcription switch toggled by noncoding RNAs. Proc Natl Acad Sci U S A 106:18049-50
Helmlinger, Dominique; Marguerat, Samuel; Villen, Judit et al. (2008) The S. pombe SAGA complex controls the switch from proliferation to sexual differentiation through the opposing roles of its subunits Gcn5 and Spt8. Genes Dev 22:3184-95
Laprade, Lisa; Rose, David; Winston, Fred (2007) Characterization of new Spt3 and TATA-binding protein mutants of Saccharomyces cerevisiae: Spt3 TBP allele-specific interactions and bypass of Spt8. Genetics 177:2007-17
Hickman, Mark J; Winston, Fred (2007) Heme levels switch the function of Hap1 of Saccharomyces cerevisiae between transcriptional activator and transcriptional repressor. Mol Cell Biol 27:7414-24

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