Probing Zn2+-binding effects on the zinc-ribbon domain of human general transcription factor TFIIB

Mahua Ghosh, Laura M. Elsby, Tapas K. Mal, Jane M. Gooding, Stefan G.E. Roberts, Mitsuhiko Ikura

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The general transcription factor, TFIIB, plays an important role in the assembly of the pre-initiation complex. The N-terminal domain (NTD) of TFIIB contains a zinc-ribbon motif, which is responsible for the recruitment of RNA polymerase II and TFIIF to the core promoter region. Although zinc-ribbon motif structures of eukaryotic and archaeal TFIIBs have been reported previously, the structural role of Zn2+ binding to TFIIB remains to be determined. In the present paper, we report NMR and biochemical studies of human TFIIB NTD, which characterize the structure and dynamics of the TFIIB Zn 2+-binding domain in both Zn2+-bound and -free states. The NMR data show that, whereas the backbone fold of NTD is pre-formed in the apo state, Zn2+ binding reduces backbone mobility in the β-turn (Arg28-Gly30), induces enhanced structural rigidity of the charged-cluster domain in the central linker region of TFIIB and appends a positive surface charge within the Zn2+-binding site. V8 protease-sensitivity assays of full-length TFIIB support the Zn2+-dependent structural changes. These structural effects of Zn2+ binding on TFIIB may have a critical role in interactions with its binding partners, such as the Rpb1 subunit of RNA polymerase II.

Original languageEnglish (US)
Pages (from-to)317-324
Number of pages8
JournalBiochemical Journal
Volume378
Issue number2
DOIs
StatePublished - Mar 1 2004

Fingerprint

Transcription Factor TFIIB
General Transcription Factors
Zinc
RNA Polymerase II
Nuclear magnetic resonance
Surface charge
Genetic Promoter Regions
Rigidity
Assays
Binding Sites

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ghosh, Mahua ; Elsby, Laura M. ; Mal, Tapas K. ; Gooding, Jane M. ; Roberts, Stefan G.E. ; Ikura, Mitsuhiko. / Probing Zn2+-binding effects on the zinc-ribbon domain of human general transcription factor TFIIB. In: Biochemical Journal. 2004 ; Vol. 378, No. 2. pp. 317-324.
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Probing Zn2+-binding effects on the zinc-ribbon domain of human general transcription factor TFIIB. / Ghosh, Mahua; Elsby, Laura M.; Mal, Tapas K.; Gooding, Jane M.; Roberts, Stefan G.E.; Ikura, Mitsuhiko.

In: Biochemical Journal, Vol. 378, No. 2, 01.03.2004, p. 317-324.

Research output: Contribution to journalArticle

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