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Mammalian transcription activation domains of VP16, AP2 and CTF activate transcription in a whole cell extract from Schizosaccharomyces pombe through the SRB/mediator
Author(s) -
Tamayo Evelyn,
Bernal Giuliano,
Maldonado Edio
Publication year - 2005
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1228
Subject(s) - general transcription factor , biology , rna polymerase ii , transcription (linguistics) , transcription factor ii f , transcription factor ii d , schizosaccharomyces pombe , transcription coregulator , schizosaccharomyces , e box , microbiology and biotechnology , response element , mediator , transcription factor , transcription factor ii b , promoter , enhancer , gene expression , transcriptional regulation , genetics , gene , saccharomyces cerevisiae , linguistics , philosophy
The acidic‐rich activation domain of VP16 and the proline‐rich activation domains of human AP2 and human CTF are able to activate transcription in a whole cell extract from Schizosaccharomyces pombe, whereas the glutamine‐rich domains of Sp1 and Oct2 are unable to activate transcription in this system. Immunodepletion experiments of the whole cell extracts using specific antibodies against pombe TAF110, pombe TAF 72, pombe TBP and Srb4 shows that the activation of transcription by VP16, AP2 and CTF is through the mediator, since depletion of Srb4 inhibits activated transcription but does not inhibit basal transcription. Immunodepletion of TBP causes inhibition of both activated and basal transcription. On the other hand, immunodepletion of TAFs does not have an effect on either activated or basal transcription. Purified RNA polymerase holoenzyme is able to rescue the transcriptional activation activity of the anti‐Srb4 immunodepleted extract. Moreover, we demonstrate that the mediator is needed for basal transcription of a TATA‐less promoter. Copyright © 2005 John Wiley & Sons, Ltd.