2UYF

Single mutant F111L DntR from Burkholderia sp. strain DNT in complex with thiocyanate

2UYF の概要

• エントリーDOI: 10.2210/pdb2uyf/pdb
• 関連するPDBエントリー: 1UTB 1UTH 2UYE
• 分子名称: REGULATORY PROTEIN, THIOCYANATE ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: lttr, lysr, dna-binding, transcription, transcription regulation, transcriptional regulator
• 由来する生物種: BURKHOLDERIA CEPACIA
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 70486.55

構造登録者

Lonneborg, R.,Smirova, I.,Dian, C.,Leonard, G.A.,Mcsweeney, S.,Brzezinski, P. (登録日: 2007-04-04, 公開日: 2007-08-21, 最終更新日: 2023-12-13)

主引用文献

Lonneborg, R.,Smirova, I.,Dian, C.,Leonard, G.A.,Brzezinski, P.
In Vivo and in Vitro Investigation of Transcriptional Regulation by Dntr.
J.Mol.Biol., 372:571-, 2007

PubMed Abstract: DntR is a bacterial transcription factor that has been isolated from Burkholderia species that are able to degrade the nitro-aromatic compound 2,4-dinitrotoluene. We recently solved the X-ray crystal structure of DntR, which suggested a putative location of an inducer-binding cavity (IBC). In this study, we constructed mutants of DntR in which residues lining the proposed IBC were modified in order to identify the structural elements involved in inducer binding, to modulate the inducer binding specificity, and to investigate the mechanism of transcriptional regulation by DntR. The transcriptional activation of the reporter gene gfp induced by the wild-type and mutant DntRs was monitored by analysing whole-cell fluorescence using flow-cytometry after addition of a number of potential inducer compounds. Three of the mutant proteins (F111L; F111V/H169V and Y110S/F111V) were purified and the binding constants for several of the potential inducers to these mutants were estimated. Furthermore, crystal structures of the F111L and Y110S/F111V mutant proteins were solved and used to explain changes in the inducer binding specificity at an atomic level. A comparison of the inducing capability in the whole-cell system and binding constants for a number of potential inducers suggests a mechanism where binding of an inducer molecule is not the sole requirement for transcriptional activation. In addition, specific interactions between DntR and the inducer molecule resulting in a conformational change of the protein are needed.

PubMed: 17681542
DOI: 10.1016/J.JMB.2007.06.076

実験手法

X-RAY DIFFRACTION (2.2 Å)