Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302

Tsung Han Chou, Jared A. Delmar, Catherine C. Wright, Nitin Kumar, Abhijith Radhakrishnan, Julia K. Doh, Meredith H. Licon, Jani Reddy Bolla, Hsiang Ting Lei, Kanagalaghatta R. Rajashankar, Chih Chia Su, Georgiana Purdy, Edward W. Yu

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Mycobacterium tuberculosis is a pathogenic bacterial species, which is neither Gram positive nor Gram negative. It has a unique cell wall, making it difficult to kill and conferring resistance to antibiotics that disrupt cell wall biosynthesis. Thus, the mycobacterial cell wall is critical to the virulence of these pathogens. Recent work shows that the mycobacterial membrane protein large (MmpL) family of transporters contributes to cell wall biosynthesis by exporting fatty acids and lipidic elements of the cell wall. The expression of the Mycobacterium tuberculosis MmpL proteins is controlled by a complicated regulatory network system. Here we report crystallographic structures of two forms of the TetR-family transcriptional regulator Rv0302, which participates in regulating the expression of MmpL proteins. The structures reveal a dimeric, two-domain molecule with architecture consistent with the TetR family of regulators. Comparison of the two Rv0302 crystal structures suggests that the conformational changes leading to derepression may be due to a rigid body rotational motion within the dimer interface of the regulator. Using fluorescence polarization and electrophoretic mobility shift assays, we demonstrate the recognition of promoter and intragenic regions of multiple mmpL genes by this protein. In addition, our isothermal titration calorimetry and electrophoretic mobility shift experiments indicate that fatty acids may be the natural ligand of this regulator. Taken together, these experiments provide new perspectives on the regulation of the MmpL family of transporters.

Original languageEnglish (US)
Pages (from-to)1942-1955
Number of pages14
JournalProtein Science
Volume24
Issue number12
DOIs
StatePublished - Dec 1 2015

Fingerprint

Mycobacterium tuberculosis
Cell Wall
Crystal structure
Cells
Membrane Proteins
Electrophoretic mobility
Biosynthesis
Fatty Acids
Calorimetry
Proteins
Fluorescence Polarization
Electrophoretic Mobility Shift Assay
Pathogens
Microbial Drug Resistance
Titration
Genetic Promoter Regions
Dimers
Virulence
Assays
Experiments

Keywords

  • Mycobacterial membrane protein
  • Mycobacterial membrane protein large
  • Rv0302
  • small
  • TetR-family regulator

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Chou, T. H., Delmar, J. A., Wright, C. C., Kumar, N., Radhakrishnan, A., Doh, J. K., ... Yu, E. W. (2015). Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302. Protein Science, 24(12), 1942-1955. https://doi.org/10.1002/pro.2802

Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302. / Chou, Tsung Han; Delmar, Jared A.; Wright, Catherine C.; Kumar, Nitin; Radhakrishnan, Abhijith; Doh, Julia K.; Licon, Meredith H.; Bolla, Jani Reddy; Lei, Hsiang Ting; Rajashankar, Kanagalaghatta R.; Su, Chih Chia; Purdy, Georgiana; Yu, Edward W.

In: Protein Science, Vol. 24, No. 12, 01.12.2015, p. 1942-1955.

Research output: Contribution to journalArticle

Chou, TH, Delmar, JA, Wright, CC, Kumar, N, Radhakrishnan, A, Doh, JK, Licon, MH, Bolla, JR, Lei, HT, Rajashankar, KR, Su, CC, Purdy, G & Yu, EW 2015, 'Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302', Protein Science, vol. 24, no. 12, pp. 1942-1955. https://doi.org/10.1002/pro.2802
Chou TH, Delmar JA, Wright CC, Kumar N, Radhakrishnan A, Doh JK et al. Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302. Protein Science. 2015 Dec 1;24(12):1942-1955. https://doi.org/10.1002/pro.2802
Chou, Tsung Han ; Delmar, Jared A. ; Wright, Catherine C. ; Kumar, Nitin ; Radhakrishnan, Abhijith ; Doh, Julia K. ; Licon, Meredith H. ; Bolla, Jani Reddy ; Lei, Hsiang Ting ; Rajashankar, Kanagalaghatta R. ; Su, Chih Chia ; Purdy, Georgiana ; Yu, Edward W. / Crystal structure of the Mycobacterium tuberculosis transcriptional regulator Rv0302. In: Protein Science. 2015 ; Vol. 24, No. 12. pp. 1942-1955.
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