HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE ADVANCED SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Tanaka, A. Articles by Ohkuma, Y. Search for Related Content PubMed Articles by Tanaka, A. Articles by Ohkuma, Y.

¹ Laboratory of Gene Regulation, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan
² Graduate School of Frontier Biosciences, and
³ Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan
⁴ Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan

The human general transcription factor, TFIIE, consists of two subunits, and β. Structural analyses indicated the presence of a forkhead motif within the central region of TFIIEβ. This motif was essential for transcription and possessed a double-stranded DNA-binding activity. Protein-DNA photo-cross-linking studies indicated that TFIIEβ binds within the promoter region, adjacent to the transcription initiation site where promoter melting begins at transcription initiation. Furthermore, neither TFIIE nor the other general transcription factor TFIIH, were required for basal transcription of adenovirus major late promoter artificially pre-melted at the initiation site. These data suggest a model in which TFIIE binds to a position adjacent to the initiation site via the forkhead domain, enabling TFIIH to begin opening the promoter. Here, we used systematic point mutations to further investigate the functional roles of this domain. The mutant proteins were expressed in bacteria, purified and used to examine transcription of two different forms of template, phosphorylation of the C-terminal domain of RNA polymerase II, as well as dsDNA-binding. Taken together, our results strongly demonstrated that the primary function of the forkhead region is dsDNA-binding in transcription. In addition, we identified three positively charged lysine residues which play a key role in this function.

^aPresent address: Discovery Research Laboratories III, Pharmaceutical Research Division, Takeda Chemical Industries Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka 532-8686, Japan.

^* Correspondence: ohkumay{at}pha.u-toyama.ac.jp