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J. Biol. Chem., Vol. 276, Issue 33, 30641-30647, August 17, 2001
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From the Department of Microbiology and Cell Biology, Indian
Institute of Science, Bangalore, India 560 012
A major role in the regulation of eukaryotic
protein-coding genes is played by the gene-specific transcriptional
regulators, which recruit the RNA polymerase II holoenzyme to the
specific promoter. Several components of the mediator complex
within the holoenzyme also have been shown to affect activation of
different subsets of genes. Only recently has it been suggested that
besides the largest subunit of RNA polymerase II, smaller subunits like Rpb3 and Rpb5 may have regulatory roles in expression of specific sets
of genes. We report here, the role of Rpb4, a non-essential subunit of core RNA polymerase II, in activation of a subset of genes
in Saccharomyces cerevisiae. We have shown below that
whereas constitutive transcription is largely unaffected, activation
from various promoters tested is severely compromised in the absence of
RPB4. This activation defect can be rescued by the
overexpression of cognate activators. We have localized the region of
Rpb4 involved in activation to the C-terminal 24 amino acids. We have
also shown here that transcriptional activation by artificial
recruitment of the TATA-binding protein (TBP) to the promoter is also
defective in the absence of RPB4. Surprisingly, the
overexpression of RPB7 (the interacting partner of Rpb4)
does not rescue the activation defect of all the promoters tested,
although it rescues the activation defect of the heat shock
element-containing promoter and the temperature sensitivity associated
with RPB4 deletion. Overall, our results indicate
that Rpb4 and Rpb7 play independent roles in transcriptional regulation
of genes.
To whom correspondence should be addressed. Tel.: 91-80-309-2292;
Fax: 91-80-360-2697; E-mail: pps@mcbl.iisc.ernet.in.
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