7DWN

Crystal structure of Vibrio fischeri DarR in complex with DNA reveals the transcriptional activation mechanism of LTTR family members

7DWN の概要

• エントリーDOI: 10.2210/pdb7dwn/pdb
• 分子名称: Predicted DNA-binding transcriptional regulator, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID (3 entities in total)
• 機能のキーワード: transcriptional regulators, tetramer, dna binding protein
• 由来する生物種: Aliivibrio fischeri ES114
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 131851.56

構造登録者

Wang, W.W.,Wu, H.,He, J.H.,Yu, F. (登録日: 2021-01-17, 公開日: 2021-07-07, 最終更新日: 2023-11-29)

主引用文献

Wang, W.,Wu, H.,Xiao, Q.,Zhou, H.,Li, M.,Xu, Q.,Wang, Q.,Yu, F.,He, J.
Crystal structure details of Vibrio fischeri DarR and mutant DarR-M202I from LTTR family reveals their activation mechanism.
Int.J.Biol.Macromol., 183:2354-2363, 2021

PubMed Abstract: DarR, a novel member of the LTTR family derived from Vibrio fischeri, activates transcription in response to d-Asp and regulates the overexpression of the racD genes encoding a putative aspartate racemase, RacD. Here, the crystal structure of full-length DarR and its mutant DarR-M202I were obtained by X-ray crystallography. According to the electron density map analysis of full-length DarR, the effector binding site of DarR is occupied by 2-Morpholinoethanesulfonic acid monohydrate (MES), which could interact with amino acids in the effector binding site and stabilize the effector binding site. Furthermore, we elaborated the structure of DarR-M202I, where methionine is replaced by isoleucine resulting in overexpression of the downstream operon. By comparing DarR-MES and DarR-M202I, we found similar behavior of DarR-MES in terms of the stability of the RD active pocket and the deflection angle of the DBD. The Isothermal titration calorimetry and Gel-filtration chromatography experiments showed that only when the target DNA sequence of a particular quasi-palindromic sequence exceeds 19 bp, DarR can effectively bind to racD promoter. This study will help enhance our understanding of the mechanism in the transcriptional regulation of LTTR family transcription factors.

PubMed: 34081954
DOI: 10.1016/j.ijbiomac.2021.05.186

実験手法

X-RAY DIFFRACTION (2.32 Å)