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

Yoshiaki Ohkuma, Hideki Sumimoto, Masami Horikoshi, Robert G. Roeder

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63 Citations (Scopus)

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-α) and 34-kDa (TFIIE-β) 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-α had an independent TFIIE activity whereas TFIIE-β had no activity alone but enhanced the activity of TFIIE-α. In conjunction with gel filtration studies, which indicated a molecular mass of ≈180 kDa for the native protein, these results suggested that TFIIE is a heterotetramer containing two a and two β 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.

Original languageEnglish
Pages (from-to)9163-9167
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number23
DOIs
Publication statusPublished - 1990

All Science Journal Classification (ASJC) codes

  • General

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