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The Structure of the Transcriptional Repressor KstR in Complex with CoA Thioester Cholesterol Metabolites Sheds Light on the Regulation of Cholesterol Catabolism in Mycobacterium tuberculosis
- Source :
- Journal of Biological Chemistry. 291:7256-7266
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Cholesterol can be a major carbon source for Mycobacterium tuberculosis during infection, both at an early stage in the macrophage phagosome and later within the necrotic granuloma. KstR is a highly conserved TetR family transcriptional repressor that regulates a large set of genes responsible for cholesterol catabolism. Many genes in this regulon, including kstR, are either induced during infection or are essential for survival of M. tuberculosis in vivo. In this study, we identified two ligands for KstR, both of which are CoA thioester cholesterol metabolites with four intact steroid rings. A metabolite in which one of the rings was cleaved was not a ligand. We confirmed the ligand-protein interactions using intrinsic tryptophan fluorescence and showed that ligand binding strongly inhibited KstR-DNA binding using surface plasmon resonance (IC50 for ligand = 25 nm). Crystal structures of the ligand-free form of KstR show variability in the position of the DNA-binding domain. In contrast, structures of KstR·ligand complexes are highly similar to each other and demonstrate a position of the DNA-binding domain that is unfavorable for DNA binding. Comparison of ligand-bound and ligand-free structures identifies residues involved in ligand specificity and reveals a distinctive mechanism by which the ligand-induced conformational change mediates DNA release.
- Subjects :
- DNA, Bacterial
0301 basic medicine
Conformational change
030106 microbiology
Repressor
Plasma protein binding
Biology
Crystallography, X-Ray
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Protein structure
Bacterial Proteins
TetR
Molecular Biology
Phagosome
Mycobacterium tuberculosis
Cell Biology
Ligand (biochemistry)
Protein Structure, Tertiary
3. Good health
Repressor Proteins
Cholesterol
Regulon
chemistry
Protein Structure and Folding
Protein Binding
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 291
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....100c8f2789390db0de6cd8a577a04732