The long-term objectives of this application are to learn more about how chromatin structure controls transcription initiation in eukaryotic cells. Histones and other functions that have been shown to be important in yeast are conserved in other eukaryotes, including humans. These functions include the SNF/SWI complex, required to overcome transcriptional repression by nucleosomes. The studies will focus on three related areas that concern how RNA polymerase II transcription initiation is controlled by factors that modulate chromatin structure. In all three areas, both genetic and biochemical approaches will be taken. First, histone mutants that alter transcription in vivo will be analyzed. Previously studied histone H2A mutants will be analyzed by identifying nonhistone proteins with which this histone interacts, as well as by biochemical analysis of nucleosomes that contain this mutant histone. Mutations in genes that encode histones H2B, H3, and H4 will also be identified and analyzed. Second, the SPT4/SPT5/SPT6 complex, required for repression by nucleosomes in vivo, will be studied both in vivo and in vitro. These analyses will include defining the interaction between the SPT6 protein and histones, the purification and analysis of the SPT4/SPT5/SPT6 complex, and study of the human homologue of SPT4. Third, the SUC2 promoter, strongly controlled by the SNF/SWI complex, will be analyzed in detail to determine the cis- acting elements and the trans-acting factors that are required for SNF/SWI control. These studies should reveal important aspects of how chromatin structure controls transcription initiation in yeast. Since histones, SNF/SWI, and SPT4/SPT5/SPT6 are all conserved, these studies will be directly applicable to understanding transcription in other eukaryotes, including humans. Since altered transcription in humans has been implicated in diseases such as cancer, these studies are relevant to human disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032967-14
Application #
2021983
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1983-12-01
Project End
1999-11-30
Budget Start
1996-12-01
Budget End
1997-11-30
Support Year
14
Fiscal Year
1997
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
Doris, Stephen M; Chuang, James; Viktorovskaya, Olga et al. (2018) Spt6 Is Required for the Fidelity of Promoter Selection. Mol Cell 72:687-699.e6
Shetty, Ameet; Kallgren, Scott P; Demel, Carina et al. (2017) Spt5 Plays Vital Roles in the Control of Sense and Antisense Transcription Elongation. Mol Cell 66:77-88.e5
Winston, Fred; Koshland, Douglas (2016) Back to the Future: Mutant Hunts Are Still the Way To Go. Genetics 203:1007-10
DeGennaro, Christine M; Alver, Burak H; Marguerat, Samuel et al. (2013) Spt6 regulates intragenic and antisense transcription, nucleosome positioning, and histone modifications genome-wide in fission yeast. Mol Cell Biol 33:4779-92
Chang, Jennifer S; Winston, Fred (2013) Cell-cycle perturbations suppress the slow-growth defect of spt10? mutants in Saccharomyces cerevisiae. G3 (Bethesda) 3:573-83
Rando, Oliver J; Winston, Fred (2012) Chromatin and transcription in yeast. Genetics 190:351-87
Kiely, Christine M; Marguerat, Samuel; Garcia, Jennifer F et al. (2011) Spt6 is required for heterochromatic silencing in the fission yeast Schizosaccharomyces pombe. Mol Cell Biol 31:4193-204
Ivanovska, Iva; Jacques, Pierre-√Čtienne; Rando, Oliver J et al. (2011) Control of chromatin structure by spt6: different consequences in coding and regulatory regions. Mol Cell Biol 31:531-41
Chang, Jennifer S; Winston, Fred (2011) Spt10 and Spt21 are required for transcriptional silencing in Saccharomyces cerevisiae. Eukaryot Cell 10:118-29
Libuda, Diana E; Winston, Fred (2010) Alterations in DNA replication and histone levels promote histone gene amplification in Saccharomyces cerevisiae. Genetics 184:985-97

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