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. 1990 Dec;87(23):9163-7.
doi: 10.1073/pnas.87.23.9163.

Factors involved in specific transcription by mammalian RNA polymerase II: purification and characterization of general transcription factor TFIIE

Y Ohkuma  1 , H Sumimoto, M Horikoshi, R G Roeder
Affiliations

Factors involved in specific transcription by mammalian RNA polymerase II: purification and characterization of general transcription factor TFIIE

Y Ohkuma et al. Proc Natl Acad Sci U S A. 1990 Dec.

Abstract

Human transcription factor TFIIE, a ubiquitous factor required for transcription initiation by RNA polymerase II, was purified to homogeneity by a combination of conventional and HPLC steps. The purified TFIIE contained equimolar amounts of 57-kDa (TFIIE-alpha) and 34-kDa (TFIIE-beta) polypeptides that were judged to be functional subunits on the basis of their copurification with transcriptional activity and the recovery of activity following renaturation of polypeptides separated by reverse-phase HPLC. TFIIE-alpha had an independent TFIIE activity whereas TFIIE-beta had no activity alone but enhanced the activity of TFIIE-alpha. In conjunction with gel filtration studies, which indicated a molecular mass of approximately 180 kDa for the native protein, these results suggested that TFIIE is a heterotetramer containing two alpha and two beta polypeptides. Functional studies with the purified TFIIE demonstrated that it is a general initiation factor, required for all of the genes tested, but it failed to show any DNA-dependent ATPase activity.

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