Isolation of yeast transcription factor IIA using a functional transcription assay.
TFIIA was extensively purified from a whole-cell transcription extract from yeast. Activity was followed throughout isolation utilizing a functional transcription assay. Transcription activity was found to copurify with polypeptides of 43 and 12.5 kDa, consistent with a previous purification that utilized a TBP/DNA gel mobility shift assay (J. Ranish and S. Hahn, J. Biol. Chem. 266, 19320-19327, 1991). The Stoke's radius of the purified protein was determined by gel filtration chromatography to be 44 A under native conditions. The solution molecular weight derived from this measurement, 110 kDa, is consistent with a heterotetrameric structure of TFIIA.
Wade
P A
PA
University of Colorado Health Sciences Center, Department of Biochemistry, Biophysics and Genetics, Denver 80262.
Jaehning
J A
JA
D002850:Chromatography, Gel; D012319:RNA Polymerase II; D035561:Transcription Factor TFIIA; D014157:Transcription Factors / Q000737:chemistry* / Q000302:isolation & purification*; D014158:Transcription, Genetic
D035561:Transcription Factor TFIIA; D014157:Transcription Factors; D012319:RNA Polymerase II