Abstract

The long-range goal of this research is to understand the molecular mechanisms involved in the transcriptional regulation of a eukaryotic gene. To achieve this goal, we will analyze the control of transcription of yeast genes required for galactose utilization (the GAL genes), in which two key regulatory gene products (GAL4 and GAL80) control the inducibility of five GAL structural genes at the level of transcription. Three interrelated lines of research will be conducted. The first is focused towards identifying DNA sequences required for promoter function and proper regulation of three GAL structural genes. This involves analyzing the affects of mutations generated in vitro on GAL-lacZ fusions. The second line of research involves constructing strains which overproduce cloned GAL regulatory gene products (GAL4 and GAL80), followed by purification and characterization of these two proteins. The third line of research is directed towards developing a transcription system for transcribing yeast genes by RNA polymerase II in vitro. At all stages of the work, we plan to correlate the results of experiments performed in vivo with those performed in vitro. This will, hopefully, enable us to draw firm conclusions concerning molecular mechanisms of transcriptional control in eukaryotes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032308-03
Application #
3281024
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1983-07-01
Project End
1988-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Harvard University
Department
Type
Schools of Arts and Sciences
DUNS #
071723621
City
Cambridge
State
MA
Country
United States
Zip Code

  Publications

Berrozpe, Georgina; Bryant, Gene O; Warpinski, Katherine et al. (2017) Polycomb Responds to Low Levels of Transcription. Cell Rep 20:785-793
Wang, Xin; Bryant, Gene O; Floer, Monique et al. (2011) An effect of DNA sequence on nucleosome occupancy and removal. Nat Struct Mol Biol 18:507-9
Wang, Xin; Muratani, Masafumi; Tansey, William P et al. (2010) Proteolytic instability and the action of nonclassical transcriptional activators. Curr Biol 20:868-71
Floer, Monique; Wang, Xin; Prabhu, Vidya et al. (2010) A RSC/nucleosome complex determines chromatin architecture and facilitates activator binding. Cell 141:407-18
Cheng, Jason X; Gandolfi, Michele; Ptashne, Mark (2004) Activation of the Gal1 gene of yeast by pairs of 'non-classical' activators. Curr Biol 14:1675-9
Cheng, Jason X; Floer, Monique; Ononaji, Paul et al. (2002) Responses of four yeast genes to changes in the transcriptional machinery are determined by their promoters. Curr Biol 12:1828-32
Lu, Zhen; Ansari, Aseem Z; Lu, Xiangyang et al. (2002) A target essential for the activity of a nonacidic yeast transcriptional activator. Proc Natl Acad Sci U S A 99:8591-6
Cheng, Jason X; Nevado, Julian; Lu, Zhen et al. (2002) The TBP-inhibitory domain of TAF145 limits the effects of nonclassical transcriptional activators. Curr Biol 12:934-7
Bamdad, C (1998) The use of variable density self-assembled monolayers to probe the structure of a target molecule. Biophys J 75:1989-96
Ansari, A Z; Reece, R J; Ptashne, M (1998) A transcriptional activating region with two contrasting modes of protein interaction. Proc Natl Acad Sci U S A 95:13543-8

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