Open AccessSimple derivation of TFIID-dependent RNA polymerase II transcription systems from Schizosaccharomyces pombe and other organisms, and factors required for transcriptional activation.
Author(s) -
Peter M. Flanagan,
Raymond J. Kelleher,
Herbert Tschochner,
Michael H. Sayre,
Roger D. Kornberg
Publication year - 1992
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.89.16.7659
Subject(s) - transcription factor ii d , schizosaccharomyces pombe , rna polymerase ii , general transcription factor , biology , transcription factor ii a , schizosaccharomyces , rna polymerase , rna polymerase iii , rna polymerase ii holoenzyme , transcription (linguistics) , transcription factor ii f , microbiology and biotechnology , transcription factor ii b , polymerase , transcription preinitiation complex , saccharomyces cerevisiae , promoter , genetics , rna , yeast , dna , gene , gene expression , linguistics , philosophy
Resolution of whole cell extract through two chromatographic steps yields a single protein fraction requiring only the addition of TFIID for the initiation of transcription at RNA polymerase II promoters. This approach allows the convenient generation of RNA polymerase II transcription systems from Saccharomyces cerevisiae, human lymphocytes, and Schizosaccharomyces pombe. TFIIDs from all three organisms are interchangeable among all three systems. The S. cerevisiae and Sch. pombe systems support effects of acidic activator proteins, provided a further protein fraction from S. cerevisiae is supplied. This further fraction is distinct from the mediator of transcriptional activation described previously and represents a second component in addition to general initiation factors that may facilitate a response to acidic activators.