DprE1 Is a Vulnerable Tuberculosis Drug Target Due to Its Cell Wall Localization

Brecik, Miroslav; Centárová, Ivana; Mukherjee, Raju; Kolly, Gaëlle S.; Huszár, Stanislav; Bobovská, Adela; Kilacskova, Emoeke; Mokosova, Veronika; Svetlíková, Zuzana; Šarkan, Michal; Neres, João; Korduláková, Jana; Cole, Stewart T.; Mikušová, Katarı́na

doi:10.1021/acschembio.5b00237

Cited by 132 publications

(105 citation statements)

References 28 publications

Supporting

Mentioning

102

Contrasting

Unclassified

Order By: Relevance

“…S12). It is unclear how Rv0560c, a putative cytoplasmic enzyme, inactivates 14 before 14 targets DprE1, whose activity has been localized to the Mtb cell wall (37). There may be an uptake mechanism for 14 that delivers most of it from the extracellular medium to the cytosol before it can reach DprE1 in the periplasm, or perhaps marked increase in expression of Rv0560c results in some of the enzyme becoming periplasmic.…”

Section: Discussionmentioning

confidence: 99%

N -methylation of a bactericidal compound as a resistance mechanism in Mycobacterium tuberculosis

Warrier

Kapilashrami

Argyrou

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The rising incidence of antimicrobial resistance (AMR) makes it imperative to understand the underlying mechanisms. Mycobacterium tuberculosis (Mtb) is the single leading cause of death from a bacterial pathogen and estimated to be the leading cause of death from AMR. A pyrido-benzimidazole, 14, was reported to have potent bactericidal activity against Mtb. Here, we isolated multiple Mtb clones resistant to 14. Each had mutations in the putative DNA-binding and dimerization domains of rv2887, a gene encoding a transcriptional repressor of the MarR family. The mutations in Rv2887 led to markedly increased expression of rv0560c. We characterized Rv0560c as an S-adenosyl-L-methionine-dependent methyltransferase that N-methylates 14, abolishing its mycobactericidal activity. An Mtb strain lacking rv0560c became resistant to 14 by mutating decaprenylphosphoryl-β-d-ribose 2-oxidase (DprE1), an essential enzyme in arabinogalactan synthesis; 14 proved to be a nanomolar inhibitor of DprE1, and methylation of 14 by Rv0560c abrogated this activity. Thus, 14 joins a growing list of DprE1 inhibitors that are potently mycobactericidal. Bacterial methylation of an antibacterial agent, 14, catalyzed by Rv0560c of Mtb, is a previously unreported mechanism of AMR.

show abstract

Section: Discussionmentioning

confidence: 99%

N -methylation of a bactericidal compound as a resistance mechanism in Mycobacterium tuberculosis

Warrier

Kapilashrami

Argyrou

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Conditional knockdown studies showed that loss of dprE1 results in a strong bactericidal effect in vitro 2. Multiple covalent3 and non‐covalent4 DprE1 inhibitors have been identified and showed in vitro and in vivo activities against Mtb , further validating DprE1 as an attractive anti‐tuberculosis (TB) drug target 5…”

mentioning

confidence: 99%

Determinants of the Inhibition of DprE1 and CYP2C9 by Antitubercular Thiophenes

et al. 2017

View full text Add to dashboard Cite

Mycobacterium tuberculosis (Mtb) DprE1, an essential isomerase for the biosynthesis of the mycobacterial cell wall, is a validated target for tuberculosis (TB) drug development. Here we report the X‐ray crystal structures of DprE1 and the DprE1 resistant mutant (Y314C) in complexes with TCA1 derivatives to elucidate the molecular basis of their inhibitory activities and an unconventional resistance mechanism, which enabled us to optimize the potency of the analogs. The selected lead compound showed excellent in vitro and in vivo activities, and low risk of toxicity profile except for the inhibition of CYP2C9. A crystal structure of CYP2C9 in complex with a TCA1 analog revealed the similar interaction patterns to the DprE1–TCA1 complex. Guided by the structures, an optimized molecule was generated with differential inhibitory activities against DprE1 and CYP2C9, which provides insights for development of a clinical candidate to treat TB.

show abstract

“…However, the presence of a single negatively charged residue in the lumen of Rv3789 is most likely incompatible with the translocation of negatively charged DPA, since it has been reported that a cationic flippase lumen is required to translocate an anionic O-antigen subunit across the hydrophobic inner membrane (42). A final and definitive piece of evidence suggesting a role for Rv3789 in protein recruitment rather than as a flippase comes from our very recent study showing that the epimerization of DPR to DPA takes place in the periplasm (43). Hence, since DPA is already located in the periplasm, a DPA flippase is unnecessary.…”

Section: Discussionmentioning

confidence: 96%

GtrA Protein Rv3789 Is Required for Arabinosylation of Arabinogalactan in Mycobacterium tuberculosis

Kolly

Mukherjee

Kilacskova³

et al. 2015

J Bacteriol

Self Cite

View full text Add to dashboard Cite

Mycobacterium tuberculosis possesses a thick and highly hydrophobic cell wall principally composed of a mycolyl-arabinogalactan-peptidoglycan complex, which is critical for survival and virulence. DprE1 is a well-characterized component of decaprenylphospho-ribose epimerase, which produces decaprenyl-phospho-arabinose (DPA) for the biosynthesis of mycobacterial arabinans. Upstream of dprE1 lies rv3789, which encodes a short transmembrane protein of the GtrA family, whose members are often involved in the synthesis of cell surface polysaccharides. We demonstrate that rv3789 and dprE1 are cotranscribed from a common transcription start site situated 64 bp upstream of rv3789. Topology mapping revealed four transmembrane domains in Rv3789 and a cytoplasmic C terminus consistent with structural models built using analysis of sequence coevolution. To investigate its role, we generated an unmarked rv3789 deletion mutant in M. tuberculosis. The mutant was characterized by impaired growth and abnormal cell morphology, since the cells were shorter and more swollen than wild-type cells. This phenotype likely stems from the decreased incorporation of arabinan into arabinogalactan and was accompanied by an accumulation of DPA. A role for Rv3789 in arabinan biosynthesis was further supported by its interaction with the priming arabinosyltransferase AftA, as demonstrated by a two-hybrid approach. Taken together, the data suggest that Rv3789 does not act as a DPA flippase but, rather, recruits AftA for arabinogalactan biosynthesis. IMPORTANCEUpstream of the essential dprE1 gene, encoding a key enzyme of the decaprenyl phospho-arabinose (DPA) pathway, lies rv3789, coding for a short transmembrane protein of unknown function. In this study, we demonstrated that rv3789 and dprE1 are cotranscribed from a common transcription start site located 64 bp upstream of rv3789 in M. tuberculosis. Furthermore, the deletion of rv3789 led to a reduction in arabinan content and to an accumulation of DPA, confirming that Rv3789 plays a role in arabinan biosynthesis. Topology mapping, structural modeling, and protein interaction studies suggest that Rv3789 acts as an anchor protein recruiting AftA, the first arabinosyl transferase. This investigation provides deeper insight into the mechanism of arabinan biosynthesis in mycobacteria. M ycobacterium tuberculosis, the pathogen responsible for most cases of human tuberculosis (TB), is surrounded by a thick highly hydrophobic cell wall, which reduces permeability and confers resistance to many antibiotics. The cell wall core is composed of a covalently bound mycolyl-arabinogalactan-peptidoglycan (mAGP) complex, which is supplemented by proteins, free lipids, and immunomodulatory lipoglycans, such as lipomannan (LM) and lipoarabinomannan (LAM) (1-3). The mAGP complex and LAM are both essential for bacterial survival and virulence (4).Arabinogalactan forms a major component of the mAGP complex. C-5 of three of the 30 alternating ␤(1¡5) and ␤(1¡6) galactofuranose (Galf) residues (positions 8, 10...

show abstract

DprE1 Is a Vulnerable Tuberculosis Drug Target Due to Its Cell Wall Localization

Cited by 132 publications

References 28 publications

N -methylation of a bactericidal compound as a resistance mechanism in Mycobacterium tuberculosis

N -methylation of a bactericidal compound as a resistance mechanism in Mycobacterium tuberculosis

Determinants of the Inhibition of DprE1 and CYP2C9 by Antitubercular Thiophenes

GtrA Protein Rv3789 Is Required for Arabinosylation of Arabinogalactan in Mycobacterium tuberculosis

Contact Info

Product

Resources

About