Answered: Experiment 1 2 3 4 5 6 7 Components…

Name: The locations of the TATA box in two species of yeast, Saccharomyces p
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Description: The locations of the TATA box in two species of yeast, Saccharomyces pombe and Saccharomyces cerevisiae, differ dramatically. The TATA box of S. pombe is about 30 nucleotides upstream of the transcription start site, similar to the location in most o

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The locations of the TATA box in two species of yeast, Saccharomyces pombe and Saccharomyces cerevisiae, differ dramatically. The TATA box of S. pombe is about 30 nucleotides upstream of the transcription start site, similar to the location in most other eukaryotic cells. However, the TATA box of S. cerevisiae is 40 to 120 nucleotides upstream of the start site. To better understand what sets the start site in these organisms, researchers at Stanford University conducted a series of experiments to determine which components of the transcription apparatus of these two species could be interchanged (Y. Li et al. 1994. Science 263:805–807). In these experiments, different general transcription factors and RNA polymerases were switched in S. pombe and S. cerevisiae, and the effects of each switch on the level of RNA synthesis and on the starting point of transcription were observed. The results from one set of experiments are shown in the table below. Components cTFIIB, cTFIIE, cTFIIF, cTFIIH are general transcription factors from S. cerevisiae. Components pTFIIB, pTFIIE, pTFIIF, pTFIIH are general transcription factors from S. pombe. Components cPol II and pPol II are RNA polymerase II from S. cerevisiae and S. pombe, respectively. The table indicates whether the component was present (+) or missing (-) in each experiment. In the accompanying gel, the presence of a band indicates that RNA was produced, and the position of the band indicates whether it was the length predicted when transcription begins 30 bp downstream of the TATA box or when it begins 40 to 120 bp downstream of the TATA box.

Q.What conclusion can you draw from these data about what components determine the start site for transcription?

Experiment
1 2 3 4 5 6 7
Components
CTFIIE
CTFIIH + CTFIIF
CTFIIB + cPol II
pPol II
PTFIIB
PTFIIE + PTFIIH + PTFIIF
Transcription at
S. pombe start site
(30 bp downstream
from TATA box)
Transcription at
S. cerevisiae start site
(40-120 bp downstream
from TATA box)
Gel

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Experiment 1 2 3 4 5 6 7 Components CTFIIE CTFIIH + CTFIIF CTFIIB + cPol II pPol II PTFIIB PTFIIE + PTFIIH + PTFIIF Transcription at S. pombe start site (30 bp downstream from TATA box) Transcription at S. cerevisiae start site (40-120 bp downstream from TATA box) Gel