PgmNr Y482: Promoter scanning during transcription initiation in Saccharomyces cerevisiae: Pol II in the “shooting gallery”.

Authors:
Craig Kaplan


Institutes
Texas A&M University, College Station, TX.


Keyword: Transcription

Abstract:

Transcription initiation is the first step in gene expression. RNA Polymerase II (Pol II) is recruited to promoter DNA by activators, and together with core promoter elements and general transcription factors (GTFs), Pol II initiates transcription. Where Pol II initiates and how efficiently it does so is determined by the biochemical properties of the transcription machinery, and how it interacts with promoter architecture. In Saccharomyces cerevisiae, Pol II finds transcription start sites (TSSs) by scanning a promoter in a directional fashion from upstream to downstream. As in higher eukaryotes, many TSSs are used at any individual promoter. We wish to understand the mechanisms by which TSSs are identified, and how TSS usage and overall efficiency are shaped by promoter architecture (promoter DNA sequence, the spatial relationship between promoter elements, and chromatin structure). To this end, we have used alteration of Pol II and GTF activities to perturb the initiation process. We find that alteration of TFIIH activity (SSL2-encoded subunit) affects TSS selection differently than alteration of Pol II (large subunit, encoded by RPO21), TFIIB (SUA7), or TFIIF (TFG2-encoded subunit) activity. Furthermore, we find that promoter scanning appears to operate at all yeast promoters, regardless of promoter class. However, promoter scanning appears to be differentially affected by promoter architecture. We integrate our observations with previous data into a model for Pol II initiation in yeast – the “shooting gallery”. In this model, Pol II catalytic activity, and the rate and processivity of the engine for Pol II scanning, TFIIH, determine the distribution of TSSs and their usage levels together with promoter sequence.



Yeast Database Genetic Index
1. gene symbol: RPO21; systematic name: YDL140C
2. gene symbol: TFG2; systematic name: YGR005C
3. gene symbol: SUA7; systematic name: YPR086W
4. gene symbol: SSL2; systematic name: YIL143C