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CD Spectroscopy to Study DNA-Protein Interactions
Published on: February 10, 2022
DNA Topoisomerases Are Required for Preinitiation Complex Assembly during GAL Gene Activation
Morten Roedgaard1, Jacob Fredsoe1, Jakob Madsen Pedersen1
1Laboratory of Genome Research, Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, DK-8000, Aarhus C, Denmark.
Abstract:
To investigate the importance of topoisomerases for transcription of the galactose induced genes, we have studied the expression of GAL1, GAL2, GAL7 and GAL10 in Saccharomyces cerevisiae cells deficient for topoisomerases I and II. We find that topoisomerases are required for transcriptional activation of the GAL genes, but are dispensable for ongoing transcription, eliminating a role of the enzymes in transcriptional elongation. Furthermore, we demonstrate that promoter chromatin remodeling of the GAL genes is unaffected in the topoisomerase deficient strain. However, the cells fail to successfully recruit RNA polymerase II due to an inability of the TATA-binding protein (TBP) to bind to the TATA box in these promoters. We therefore argue that topoisomerases are required for accurate assembly of the preinitiation complex at the promoters of the GAL genes.
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