Structure determination of the minimal complex between Tfb5 and Tfb2, two subunits of the yeast transcription/DNA-repair factor TFIIH: a retrospective study - Archive ouverte HAL Access content directly
Journal Articles Acta crystallographica Section D : Structural biology [1993-...] Year : 2010

Structure determination of the minimal complex between Tfb5 and Tfb2, two subunits of the yeast transcription/DNA-repair factor TFIIH: a retrospective study

Abstract

Tfb5 interacts with the Tfb2 subunit of the general transcription factor TFIIH to ensure efficient nucleotide-excision repair in eukaryotes. The crystal structure of the complex between Tfb5 and the C-terminal region of Tfb2 (Tfb2C) from Saccharomyces cerevisiae has recently been reported. Here, the structure-determination process is described as a case study. Although crystals were obtained readily, it was not possible to determine experimental phases from a first crystal form (Tfb2 412–513 –Tfb5 2–72 ) that diffracted to 2.6 Å resolution. Shortening of the Tfb2C from its N-terminus was decisive and modified the crystal packing, leading to a second crystal form (Tfb2 435–513 –Tfb5 2–72 ). These crystals diffracted to 1.7 Å resolution with excellent mosaicity and allowed structure determination by conventional approaches using heavy atoms. The refined structure from the second crystal form was used to solve the structure of the first crystal form by molecular replacement. Comparison of the two structures revealed that the N-terminal region of Tfb2C and (to a lesser extent) the C-terminal region of Tfb5 contributed to the crystal packing. A detailed analysis illustrates how variation in domain boundaries influences crystal packing and quality.

Dates and versions

hal-03610116 , version 1 (16-03-2022)

Identifiers

Cite

Denis Kainov, Vincent Cura, Marc Vitorino, Helène Nierengarten, Pierre Poussin, et al.. Structure determination of the minimal complex between Tfb5 and Tfb2, two subunits of the yeast transcription/DNA-repair factor TFIIH: a retrospective study. Acta crystallographica Section D : Structural biology [1993-..], 2010, 66 (7), pp.745-755. ⟨10.1107/S0907444910009844⟩. ⟨hal-03610116⟩
6 View
0 Download

Altmetric

Share

Gmail Facebook X LinkedIn More