5TMX

Solution Structure of SinI, antagonist to the master biofilm-regulator SinR in Bacillus subtilis

5TMX の概要

• エントリーDOI: 10.2210/pdb5tmx/pdb
• 関連するPDBエントリー: 5TN0 5TN2
• NMR情報: BMRB: 30192
• 分子名称: Protein SinI (1 entity in total)
• 機能のキーワード: sporulation, repressor, biofilm, transcription regulator
• 由来する生物種: Bacillus subtilis (strain 168)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 14368.30

構造登録者

Draughn, G.L.,Bobay, B.G.,Stowe, S.D.,Thompson, R.J.,Cavanagh, J. (登録日: 2016-10-13, 公開日: 2017-10-25, 最終更新日: 2024-05-15)

主引用文献

Milton, M.E.,Draughn, G.L.,Bobay, B.G.,Stowe, S.D.,Olson, A.L.,Feldmann, E.A.,Thompson, R.J.,Myers, K.H.,Santoro, M.T.,Kearns, D.B.,Cavanagh, J.
The Solution Structures and Interaction of SinR and SinI: Elucidating the Mechanism of Action of the Master Regulator Switch for Biofilm Formation in Bacillus subtilis.
J.Mol.Biol., 2019

PubMed Abstract: Bacteria have developed numerous protection strategies to ensure survival in harsh environments, with perhaps the most robust method being the formation of a protective biofilm. In biofilms, bacterial cells are embedded within a matrix that is composed of a complex mixture of polysaccharides, proteins, and DNA. The gram-positive bacterium Bacillus subtilis has become a model organism for studying regulatory networks directing biofilm formation. The phenotypic transition from a planktonic to biofilm state is regulated by the activity of the transcriptional repressor, SinR, and its inactivation by its primary antagonist, SinI. In this work, we present the first full-length structural model of tetrameric SinR using a hybrid approach combining high-resolution solution nuclear magnetic resonance (NMR), chemical cross-linking, mass spectrometry, and molecular docking. We also present the solution NMR structure of the antagonist SinI dimer and probe the mechanism behind the SinR-SinI interaction using a combination of biochemical and biophysical techniques. As a result of these findings, we propose that SinI utilizes a residue replacement mechanism to block SinR multimerization, resulting in diminished DNA binding and concomitant decreased repressor activity. Finally, we provide an evidence-based mechanism that confirms how disruption of the SinR tetramer by SinI regulates gene expression.

PubMed: 31493408
DOI: 10.1016/j.jmb.2019.08.019

実験手法

SOLUTION NMR