Interdependent interactions between TFIIB, TATA binding protein, and DNA.

Abstract:

:Temperature-sensitive mutants of TFIIB that are defective for essential interactions were isolated. One mutation (G204D) results in disruption of a protein-protein contact between TFIIB and TATA binding protein (TBP), while the other (K272I) disrupts an interaction between TFIIB and DNA. The TBP gene was mutagenized, and alleles that suppress the slow-growth phenotypes of the TFIIB mutants were isolated. TFIIB with the G204D mutation [TFIIB(G204D)] was suppressed by hydrophobic substitutions at lysine 239 of TBP. These changes led to increased affinity between TBP and TFIIB. TFIIB(K272I) was weakly suppressed by TBP mutants in which K239 was changed to hydrophobic residues. However, this mutant TFIIB was strongly suppressed by conservative substitutions in the DNA binding surface of TBP. Biochemical characterization showed that these TBP mutants had increased affinity for a TATA element. The TBPs with increased affinity could not suppress TFIIB(G204D), leading us to propose a two-step model for the interaction between TFIIB and the TBP-DNA complex.

journal_name

Mol Cell Biol

authors

Buratowski RM,Downs J,Buratowski S

doi

10.1128/mcb.22.24.8735-8743.2002

subject

Has Abstract

pub_date

2002-12-01 00:00:00

pages

8735-43

issue

24

eissn

0270-7306

issn

1098-5549

journal_volume

22

pub_type

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