Structural and functional characterization of Rv0792c from Mycobacterium tuberculosis: identifying small molecule inhibitors against GntR protein

Abstract

In order to adapt in host tissues, microbial pathogens regulate their gene expression through an array of transcription factors. Here, we have functionally characterized Rv0792c, a GntR homolog from M. tuberculosis. In comparison to the parental strain, ΔRv0792c mutant strain of M. tuberculosis was compromised for survival upon exposure to oxidative stress, cell wall agents and infection in guinea pigs. RNA-seq analysis revealed that Rv0792c regulates the expression of genes that are involved in stress adaptation and virulence of M. tuberculosis. Solution small angle X-ray scattering (SAXS) data steered model building confirmed that the C-terminal region plays a pivotal role in dimer formation. Systematic evolution of ligands by exponential enrichment resulted in identification of ssDNA aptamers that can be used as a tool to identify small molecule inhibitors targeting Rv0792c. Using SELEX and SAXS data based modelling, we identified residues essential for the DNA binding activity of Rv0792c and I-OMe-Tyrphostin as an inhibitor of Rv0792c aptamer binding activity. Taken together, we provide a detailed shape-function characterization of GntR family of transcription factors from M. tuberculosis. To the best of our knowledge, this is the first study that has resulted in the identification of small molecule inhibitors against GntR family of transcription factors from bacterial pathogens.

Transcriptional regulation and drug resistance in Mycobacterium tuberculosis

“…Deletion of rv0792c impaired the ability of M. tuberculosis to infect guinea pigs, however no difference was observed in survival during exposure to INH, RIF, or LEV (Chauhan et al, 2021). A role in DR for the remaining GntR family members has yet to be identified, although some of them have been found upregulated in drug resistant isolates.…”

“…In M. tuberculosis there are at least eight putative GntR-like proteins: Rv0043c, Rv0165c (Mce1R), Rv0494, Rv0586 (Mce2R), Rv0792c (MoyR), Rv1152, Rv3060c, and Rv3575c (Vindal et al, 2007;Chauhan et al, 2021). Among them, Rv0494 has an ortholog in M. smegmatis (Ms2173).…”