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. 2011:2011:206290.
doi: 10.4061/2011/206290. Epub 2011 Oct 12.

The Transition of Poised RNA Polymerase II to an Actively Elongating State Is a "Complex" Affair

Marie N Yearling  1 , Catherine A Radebaugh, Laurie A Stargell
Affiliations

The Transition of Poised RNA Polymerase II to an Actively Elongating State Is a "Complex" Affair

Marie N Yearling et al. Genet Res Int. 2011.

Abstract

The initial discovery of the occupancy of RNA polymerase II at certain genes prior to their transcriptional activation occurred a quarter century ago in Drosophila. The preloading of these poised complexes in this inactive state is now apparent in many different organisms across the evolutionary spectrum and occurs at a broad and diverse set of genes. In this paper, we discuss the genetic and biochemical efforts in S. cerevisiae to describe the conversion of these poised transcription complexes to the active state for productive elongation. The accumulated evidence demonstrates that a multitude of coactivators and chromatin remodeling complexes are essential for this transition.

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Figures

Figure 1
Figure 1
The poised CYC1 promoter contains preloaded transcription components. (a) Prior to activation, the preloaded CYC1 promoter contains TATA binding protein (TBP), RNA polymerase II (RNAPII), the core TFIIH complex (TFIIHc), Spt-Ada-GCN5 acetyltransferase (SAGA), and the transcription factor Spn1. The CTD, shown by the hashed line trailing RNAPII exhibits serine 5 phosphorylation potentially on multiple repeats (denoted by “n”). These components occupy the promoter prior to high levels of transcriptional output. (b) The occupancy of the preloaded factors is maintained under induced conditions, and Mediator, Spt6, and Swi/Snf are recruited, leading to an increase in transcriptional output (indicated by the arrow).
Figure 2
Figure 2
Mutating SPN1, which results in loss of Spn1 and constitutive recruitment of Swi/Snf to the promoter, does not bypass the requirement for SAGA or Mediator. CYC1 transcript levels were analyzed before and two or four hours after induction in ethanol using an S1 nuclease protection assay [23] with RNA isolated from wild-type, ada1Δ or med20Δ strains. Each strain harbors either a wild-type (WT) or mutant (MT) form of SPN1. Similar results were obtained for other SAGA and Mediator deletion strains, including gcn5Δ, spt7Δ, spt8Δ, spt20Δ, med5Δ, med15Δ, and med18Δ (data not shown).
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