4W97: Structure Of Ketosteroid Transcriptional Regulator Kstr2 Of Mycobacterium Tuberculosis

Catabolism of host cholesterol is critical to the virulence of Mycobacterium tuberculosis and is a potential target for novel therapeutics. KstR2, a TetR family repressor (TFR), regulates the expression of 15 genes encoding enzymes that catabolize the last half of the cholesterol molecule, represented by 3aalpha-H-4alpha(3'-propanoate)-7abeta-methylhexahydro-1,5-indane-dione (HIP). Binding of KstR2 to its operator sequences is relieved upon binding of HIP-CoA. A 1.6-A resolution crystal structure of the KstR2Mtb.HIP-CoA complex reveals that the KstR2Mtb dimer accommodates two molecules of HIP-CoA. Each ligand binds in an elongated cleft spanning the dimerization interface such that the HIP and CoA moieties interact with different KstR2Mtb protomers. In isothermal titration calorimetry studies, the dimer bound 2 eq of HIP-CoA with high affinity (Kd = 80 +/- 10 nm) but bound neither HIP nor CoASH. Substitution of Arg-162 or Trp-166, residues that interact, respectively, with the diphosphate and HIP moieties of HIP-CoA, dramatically decreased the affinity of KstR2Mtb for HIP-CoA but not for its operator sequence. The variant of R162M that decreased the affinity for HIP-CoA (DeltaDeltaG = 13 kJ mol(-1)) is consistent with the loss of three hydrogen bonds as indicated in the structural data. A 24-bp operator sequence bound two dimers of KstR2. Structural comparisons with a ligand-free rhodococcal homologue and a DNA-bound homologue suggest that HIP-CoA induces conformational changes of the DNA-binding domains of the dimer that preclude their proper positioning in the major groove of DNA. The results provide insight into KstR2-mediated regulation of expression of steroid catabolic genes and the determinants of ligand binding in TFRs.
PDB ID: 4W97Download
MMDB ID: 125130
PDB Deposition Date: 2014/8/27
Updated in MMDB: 2014/12
Experimental Method:
x-ray diffraction
Resolution: 1.6  Å
Source Organism:
Similar Structures:
Biological Unit for 4W97: dimeric; determined by author and by software (PISA)
Molecular Components in 4W97
Label Count Molecule
Proteins (2 molecules)
Hth-type Transcriptional Repressor Kstr2(Gene symbol: Rv3557c)
Molecule annotation
Chemicals (10 molecules)
* Click molecule labels to explore molecular sequence information.

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