The regulatory circuit that acts upon the GAL genes of the yeast S. cerevisiae is well defined genetically. Of central interest is the mechanism by which the GAL4-encoded regulatory protein activates transcription of the GAL genes. This protein binds to sequences located upstream of each gene that is regulates; the events that occur subsequent to DNA binding that lead to activation of transcription are unknown. Also poorly understood are the factors responsible for catabolite repression of the GAL genes caused by growth on glucose. The long-term goal of the proposed research is to learn how proteins interact with each other and with the GAL control regions to regulate transcription. We will focus on three types of proteins: 1) the GAL4-encoded protein, 2) proteins that interact with the GAL4 protein, and 3) proteins responsible for catabolite repression of the GAL genes.
The specific aims are: 1) To locate functional regions of the GAL4-encoded protein by identifying mutations that affect the DNA binding, transcription activation, and other functions of this protein. Because the location of functional regions in this large protein are poorly defined, several in vivo will then be carried out on more precisely defined regions. 2) To identify proteins that interact with the GAL4 protein. The GAL4 protein likely activates transcription by interacting with proteins that act close to the transcription initiation site, but genes encoding such proteins have not been transcription initiation site, but genes encoding such proteins have not been identified. The genetic selections proposed here are aimed at identification of these genes. 3) To elucidate the mechanism of catabolite repression of the GAL genes by isolating and characterizing mutants defective in this regulatory circuit, using selection schemes specifically developed for this purpose. 4) To develop a genetic method for the analysis of DNA binding proteins. These methods will be applied to the study of GAL4 protein, as well as other DNA binding proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032540-06
Application #
3281471
Study Section
Genetics Study Section (GEN)
Project Start
1983-08-01
Project End
1992-03-31
Budget Start
1989-04-01
Budget End
1990-03-31
Support Year
6
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Snowdon, Chris; Johnston, Mark (2016) A novel role for yeast casein kinases in glucose sensing and signaling. Mol Biol Cell 27:3369-3375
Simpson-Lavy, Kobi J; Bronstein, Alex; Kupiec, Martin et al. (2015) Cross-Talk between Carbon Metabolism and the DNA Damage Response in S. cerevisiae. Cell Rep 12:1865-75
Simpson-Lavy, Kobi J; Johnston, Mark (2013) SUMOylation regulates the SNF1 protein kinase. Proc Natl Acad Sci U S A 110:17432-7
Libkind, Diego; Hittinger, Chris Todd; Valerio, Elisabete et al. (2011) Microbe domestication and the identification of the wild genetic stock of lager-brewing yeast. Proc Natl Acad Sci U S A 108:14539-44
Kuttykrishnan, Sooraj; Sabina, Jeffrey; Langton, Laura L et al. (2010) A quantitative model of glucose signaling in yeast reveals an incoherent feed forward loop leading to a specific, transient pulse of transcription. Proc Natl Acad Sci U S A 107:16743-8
Hittinger, Chris Todd; Gonçalves, Paula; Sampaio, José Paulo et al. (2010) Remarkably ancient balanced polymorphisms in a multi-locus gene network. Nature 464:54-8
Sabina, Jeffrey; Johnston, Mark (2009) Asymmetric signal transduction through paralogs that comprise a genetic switch for sugar sensing in Saccharomyces cerevisiae. J Biol Chem 284:29635-43
Brown, Victoria; Sabina, Jeffrey; Johnston, Mark (2009) Specialized sugar sensing in diverse fungi. Curr Biol 19:436-41
Kim, Jeong-Ho; Johnston, Mark (2006) Two glucose-sensing pathways converge on Rgt1 to regulate expression of glucose transporter genes in Saccharomyces cerevisiae. J Biol Chem 281:26144-9
Kim, Jeong-Ho; Brachet, Valerie; Moriya, Hisao et al. (2006) Integration of transcriptional and posttranslational regulation in a glucose signal transduction pathway in Saccharomyces cerevisiae. Eukaryot Cell 5:167-73

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