Lysyl-tRNA synthetase, a target for urgently needed M. tuberculosis drugs

Green, Simon R.; Davis, Susan H.; Damerow, Sebastian; Engelhart, Curtis A.; Mathieson, Michael; Baragaña, Beatriz; Robinson, David A.; Tamjar, J.; Dawson, Alice; Tamaki, Fábio K.; Buchanan, Kirsteen; Post, John; Dowers, Karen; Jansen, Chimed; Zuccotto, Fabio; Gilbert, Ian H.; Epemolu, Ola; Riley, Jennifer; Stojanovski, Laste; Osuna‐Cabello, Maria; Pérez-Herrán, Esther; Rebollo, María José; López, Laura Guijarro; Casado, Patricia; Camino, Isabel; Kim, Heather C.; Bean, James; Nahiyaan, Navid; Rhee, Kyu Y.; Wang, Qinglan; Tan, Vee Y.; Boshoff, Helena I.; Converse, Paul J.; Li, Si-Yang; Chang, Yong S.; Fotouhi, Nader; Upton, Anna M.; Nuermberger, Eric; Schnappinger, Dirk; Read, Kevin D.; Encinas, Lourdes; Bates, Robert H.; Wyatt, Paul G.; Cleghorn, Laura A. T.

doi:10.1038/s41467-022-33736-5

Cited by 15 publications

(9 citation statements)

References 39 publications

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“…Drugs that inhibit aminoacyl tRNA synthetases have seen clinical success against malaria (halofuginone) and certain skin infections (mupirocin), and efforts are under way to develop similar inhibitors for TB therapeutics. We tested a candidate lysine tRNA synthetase inhibitor (compound 7 [ 29 ]) and found a roughly 4-fold decrease in the MIC with Pth knockdowns compared to the uninduced controls ( Fig. 5 ).…”

Section: Resultsmentioning

confidence: 99%

Peptidyl tRNA Hydrolase Is Required for Robust Prolyl-tRNA Turnover in Mycobacterium tuberculosis

Tomasi

Schweber

Kimura

et al. 2023

mBio

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Section: Resultsmentioning

confidence: 99%

Peptidyl tRNA Hydrolase Is Required for Robust Prolyl-tRNA Turnover in Mycobacterium tuberculosis

Tomasi

Schweber

Kimura

et al. 2023

mBio

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“… Mtb growth in the presence a candidate lysine tRNA synthetase inhibitors [31] is plotted across drug concentrations as determined by fluorescence in an Alamar blue assay. Pth CRISPRi strains were grown to mid-log and diluted to an OD 600 of 0.001 and plated in serial dilutions of antibiotic in 96-well plates.…”

Section: Resultsmentioning

confidence: 99%

Peptidyl tRNA hydrolase is required for robust prolyl-tRNA turnover inMycobacterium tuberculosis

Tomasi

Schweber

Kimura

et al. 2022

Preprint

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Enzymes involved in rescuing stalled ribosomes and recycling translation machinery are ubiquitous in bacteria and required for growth. Peptidyl tRNA drop-off is a type of abortive translation that results in the release of a truncated peptide that is still bound to tRNA (peptidyl tRNA) into the cytoplasm. Peptidyl tRNA hydrolase (Pth) recycles the released tRNA by cleaving off the unfinished peptide and is essential in most bacterial species. We developed a sequencing-based strategy called Cu-tRNAseq to study the physiological role of Pth in Mycobacterium tuberculosis (Mtb). While most peptidyl tRNA species accumulated in a strain with impaired Pth expression, peptidyl prolyl-tRNA was particularly enriched, suggesting that Pth is required for robust peptidyl prolyl-tRNA turnover. Reducing Pth levels increased Mtb's susceptibility to tRNA synthetase inhibitors that are in development to treat tuberculosis (TB) and rendered this pathogen highly susceptible to macrolides, drugs that are ordinarily ineffective against Mtb. Collectively, our findings reveal the potency of Cu-tRNAseq for profiling peptidyl tRNAs and suggest that targeting Pth would open new therapeutic approaches for TB.

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“…Chromosomal DNA was isolated as previously described. 39 Whole genome sequencing (WGS) libraries were prepared using the KAPA HyperPlus Kit (Roche catalog number 07962380001, 07962401001, and/or 07962428001) according to manufacturer's instructions with the following modifications: in Step 1. 40 Bam files were sorted and merged using samtools.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Inhibitors of the Thioesterase Activity of Mycobacterium tuberculosis Pks13 Discovered Using DNA-Encoded Chemical Library Screening

Krieger,

Yalamanchili,

Dickson

et al. 2024

ACS Infect. Dis.

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DNA-encoded chemical library (DEL) technology provides a time-and cost-efficient method to simultaneously screen billions of compounds for their affinity to a protein target of interest. Here we report its use to identify a novel chemical series of inhibitors of the thioesterase activity of polyketide synthase 13 (Pks13) from Mycobacterium tuberculosis (Mtb). We present three chemically distinct series of inhibitors along with their enzymatic and Mtb whole cell potency, the measure of on-target activity in cells, and the crystal structures of inhibitor-enzyme complexes illuminating their interactions with the active site of the enzyme. One of these inhibitors showed a favorable pharmacokinetic profile and demonstrated efficacy in an acute mouse model of tuberculosis (TB) infection. These findings and assay developments will aid in the advancement of TB drug discovery.

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Lysyl-tRNA synthetase, a target for urgently needed M. tuberculosis drugs

Cited by 15 publications

References 39 publications

Peptidyl tRNA Hydrolase Is Required for Robust Prolyl-tRNA Turnover in Mycobacterium tuberculosis

Peptidyl tRNA Hydrolase Is Required for Robust Prolyl-tRNA Turnover in Mycobacterium tuberculosis

Peptidyl tRNA hydrolase is required for robust prolyl-tRNA turnover inMycobacterium tuberculosis

Inhibitors of the Thioesterase Activity of Mycobacterium tuberculosis Pks13 Discovered Using DNA-Encoded Chemical Library Screening

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