The head module of Mediator directs activation of preloaded RNAPII in vivo

Nucleic Acids Res. 2013 Dec;41(22):10124-34. doi: 10.1093/nar/gkt796. Epub 2013 Sep 4.

Abstract

The successful synthesis of a transcript by RNA polymerase II (RNAPII) is a multistage process with distinct rate-limiting steps that can vary depending on the particular gene. A growing number of genes in a variety of organisms are regulated at steps after the recruitment of RNAPII. The best-characterized Saccharomyces cerevisiae gene regulated in this manner is CYC1. This gene has high occupancy of RNAPII under non-inducing conditions, defining it as a poised gene. Here, we find that subunits of the head module of Mediator, Med18 and Med20, and Med19 are required for activation of transcription at the CYC1 promoter in response to environmental cues. These subunits of Mediator are required at the preloaded promoter for normal levels of recruitment and activity of the general transcription factor TFIIH. Strikingly, these Mediator components are dispensable for activation by the same activator at a different gene, which lacks a preloaded polymerase in the promoter region. Based on these results and other studies, we speculate that Mediator plays an essential role in triggering an inactive polymerase at CYC1 into a productively elongating form.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon / metabolism
  • Cytochromes c / genetics*
  • Cytochromes c / metabolism
  • Gene Deletion
  • Gene Expression Regulation, Fungal*
  • Mediator Complex / genetics
  • Mediator Complex / metabolism
  • Mediator Complex / physiology*
  • Oxidative Stress / genetics
  • Promoter Regions, Genetic
  • RNA Polymerase II / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*
  • Transcription Factors / physiology

Substances

  • CYC1 protein, S cerevisiae
  • Mediator Complex
  • ROX3 protein, S cerevisiae
  • SRB2 protein, S cerevisiae
  • SRB5 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • YAP1 protein, S cerevisiae
  • Carbon
  • Cytochromes c
  • RNA Polymerase II