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Investigation Of The Behavior Of The Gal4 Inhibitor Gal80 Of The GAL Genetic Switch In The Yeast Saccharomyces Cerevisiae

Goswami, Sudip
http://rave.ohiolink.edu/etdc/view?acc_num=osu1408542557

Abstract Details

2014, Doctor of Philosophy, Ohio State University, Molecular Genetics.
The DNA-binding transcriptional activator Gal4 and its regulators Gal80 and Gal3 constitute a galactose-responsive switch for the GAL genes of Saccharomyces cerevisiae. Gal4 binds to upstream activation sequences or UASGAL sites on GAL gene promoters as a dimer both in the absence and presence of galactose. In the absence of galactose, a Gal80 dimer binds to and masks the Gal4 activation domain, inhibiting its activity. In the presence of galactose, Gal3 interacts with Gal80 and relieves Gal80’s inhibition of Gal4 activity allowing rapid induction of expression of GAL genes. In the first part of this work (Chapter 2) in-vitro chemical crosslinking coupled with SDS PAGE and native PAGE analysis were employed to show that the presence of Gal3 that can interact with Gal80 impairs Gal80 self association. In addition, live cell spinning disk confocal imaging showed that dissipation of newly discovered Gal80-2mYFP/2GFP clusters in galactose is dependent on Gal3’s ability to interact with Gal80. In the second part (Chapter 3), extensive analysis of Gal80 clusters was carried out which showed that these clusters associate strongly with the GAL1-10-7 locus and this association is dependent on the presence of the UASGAL sites at this locus. Moreover, the formation of Gal80 clusters is dependent on the presence of Gal4. Additionally, it was discovered that Gal4-2GFP/2mYFP makes intranuclear foci, which unlike Gal80-2mYFP/2GFP clusters do not dissipate in galactose and their formation is not dependent on the presence of Gal80. Similar to Gal80 clusters, these Gal4 foci associate strongly with the GAL1-10-7 locus. These evidences suggest that large molecular assemblies of Gal80 and Gal4 associate with the UASGAL sites of the GAL1-10-7 locus, which raises some interesting possibilities as to how this genetic switch is regulated.
James Hopper (Advisor)
Stephen Osmani (Committee Member)
Hay-Oak Park (Committee Member)
JIan-Qiu Wu (Committee Member)
181 p.
Cellular Biology
GAL genetic switch

Recommended Citations

Citations

  • Goswami, S. (2014). Investigation Of The Behavior Of The Gal4 Inhibitor Gal80 Of The GAL Genetic Switch In The Yeast Saccharomyces Cerevisiae [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1408542557

    APA Style (7th edition)

  • Goswami, Sudip. Investigation Of The Behavior Of The Gal4 Inhibitor Gal80 Of The GAL Genetic Switch In The Yeast Saccharomyces Cerevisiae . 2014. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1408542557.

    MLA Style (8th edition)

  • Goswami, Sudip. "Investigation Of The Behavior Of The Gal4 Inhibitor Gal80 Of The GAL Genetic Switch In The Yeast Saccharomyces Cerevisiae ." Doctoral dissertation, Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1408542557

    Chicago Manual of Style (17th edition)

osu1408542557
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This open access ETD is published by The Ohio State University and OhioLINK.