DNA Topoisomerases Are Required for Preinitiation Complex Assembly during GAL Gene Activation

PLoS One. 2015 Jul 14;10(7):e0132739. doi: 10.1371/journal.pone.0132739. eCollection 2015.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin Assembly and Disassembly
  • DNA Topoisomerases / metabolism*
  • Galactokinase / genetics
  • Galactose / metabolism*
  • Genes, Fungal*
  • Monosaccharide Transport Proteins / genetics
  • Promoter Regions, Genetic
  • RNA Polymerase II / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • TATA-Box Binding Protein / metabolism
  • Trans-Activators / genetics
  • Transcriptional Activation

Substances

  • GAL10 protein, S cerevisiae
  • GAL2 protein, S cerevisiae
  • Monosaccharide Transport Proteins
  • Saccharomyces cerevisiae Proteins
  • TATA-Box Binding Protein
  • Trans-Activators
  • GAL1 protein, S cerevisiae
  • Galactokinase
  • RNA Polymerase II
  • DNA Topoisomerases
  • Galactose

Grants and funding

This work was supported by the Danish Natural Science Research Council, the Novo Nordisk Foundation, the Karen Elise Jensen Foundation, the Gangsted Foundation, the Simon Fougner Hartmanns Foundation, the Dagmar Marshalls Foundation, the Astrid Thaysens Foundation, and the Villum Kann Rasmussen Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.