Quantitation of the RNA polymerase II transcription machinery in yeast.
TAP tags and dot blot analysis have been used to measure the amounts of RNA polymerase II transcription proteins in crude yeast extracts. The measurements showed comparable amounts of RNA polymerase II, TFIIE, and TFIIF, lower levels of TBP and TFIIB, and still lower levels of Mediator and TFIIH. These findings are consistent with the presumed roles of the transcription proteins, but do not support the idea of their recruitment in a single large complex to RNA polymerase II promoters. The approach employed here can be readily extended to quantitative analysis of the entire yeast proteome.
Borggrefe
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Department of Structural Biology, Stanford University School of Medicine, Stanford, California 9430.
Davis
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Bareket-Samish
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Kornberg
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RD
D002455:Cell Division; D004591:Electrophoresis, Polyacrylamide Gel; D005821:Genetic Techniques; D005982:Glutathione Transferase / Q000378:metabolism; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000378:metabolism; D011485:Protein Binding; D012319:RNA Polymerase II / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D035581:Transcription Factor TFIIB; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000096:biosynthesis; D019473:Transcription Factors, TFII / Q000096:biosynthesis; D014158:Transcription, Genetic
D007074:Immunoglobulin G; D035581:Transcription Factor TFIIB; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D019473:Transcription Factors, TFII; C066644:transcription factor TFIIE; D005982:Glutathione Transferase; D012319:RNA Polymerase II; C056604:transcription factor TFIIF