Crystal structure of the transcriptional regulator Rv0678 of Mycobacterium tuberculosis [Protein Structure and Folding]

April 15th, 2014 by Radhakrishnan, A., Kumar, N., Wright, C. C., Chou, T.-H., Tringides, M. L., Bolla, J. R., Lei, H.-T., Rajashankar, K., Su, C.-C., Purdy, G. E., Yu, E. W.

Recent work demonstrates that the Mycobacterial membrane protein large (MmpL) transporters are dedicated to the export of mycobacterial lipids for cell wall biosynthesis. An MmpL transporter frequently works with an accessory protein, belonging to the Mycobacterial membrane protein small (MmpS) family, to transport these key virulence factors. One such efflux system in Mycobacterium tuberculosis is the MmpS5-MmpL5 transporter. The expression of MmpS5-MmpL5 is controlled by the MarR-like transcriptional regulator Rv0678, whose open reading frame is located downstream of the mmpS5-mmpL5 operon. To elucidate the structural basis of Rv0678 regulation, we have determined the crystal structure of this regulator, to 1.64 angstrom resolution, revealing a dimeric two-domain molecule with an architecture similar to members of the MarR family of transcriptional regulators. Rv0678 is distinct from other MarR regulators in that its DNA-binding and dimerization domains are clustered together. These two domains seemingly cooperate to bind an inducing ligand that we identified as 2-palmitoylglycerol, which is a fatty acid glycerol ester. The structure also suggests that the conformational change leading to substrate-mediated derepression is primarily caused by a rigid body rotational motion of the entire DNA-binding domain of the regulator towards the dimerization domain. This movement results in a conformational state that is incompatible with DNA binding. We demonstrate using electrophoretic mobility shift assays that Rv0678 binds to the mmpS5-mmpL5, mmpS4-mmpL4 and the mmpS2-mmpL2 promoters. Binding by Rv0678 was reversed upon addition of the ligand. These findings provide new insight into the mechanisms of gene regulation in the MarR family of regulators.