Crystal structure of the transcriptional regulator Rv0678 of Mycobacterium tuberculosis

Abhijith Radhakrishnan, Nitin Kumar, Catherine C. Wright, Tsung Han Chou, Marios L. Tringides, Jani Reddy Bolla, Hsiang Ting Lei, Kanagalaghatta R. Rajashankar, Chih Chia Su, Georgiana E. Purdy, Edward W. Yu

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Recent work demonstrates that the MmpL(mycobacterial membrane protein large) 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 MmpS (mycobacterial membrane protein small) family, to transport these key virulence factors. One such efflux system in Mycobacterium tuberculosis is the MmpS5-MmpL5 transporter. The expression of MmpS5-MmpL5is controlled by the MarR-like transcriptional regulator Rv0678, whose open reading frame is located down stream of the mmpS5-mmpL5operon. To elucidate the structural basis of Rv0678 regulation, we have determined the crystal structure of this regulator, to 1.64Ȧ 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-stearoylglycerol, 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 toward 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 the addition of the ligand. These findings provide new insight into the mechanisms of gene regulation in the MarR family of regulators.

Original languageEnglish (US)
Pages (from-to)16526-16540
Number of pages15
JournalJournal of Biological Chemistry
Volume289
Issue number23
DOIs
StatePublished - Jun 6 2014

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Radhakrishnan, A., Kumar, N., Wright, C. C., Chou, T. H., Tringides, M. L., Bolla, J. R., Lei, H. T., Rajashankar, K. R., Su, C. C., Purdy, G. E., & Yu, E. W. (2014). Crystal structure of the transcriptional regulator Rv0678 of Mycobacterium tuberculosis. Journal of Biological Chemistry, 289(23), 16526-16540. https://doi.org/10.1074/jbc.M113.538959