DNA-Dependent Binding of Nargenicin to DnaE1 Inhibits Replication in <i>Mycobacterium tuberculosis</i>

Chengalroyen, Melissa; Mason, Mandy K; Borsellini, Alessandro; Tassoni, R.; Abrahams, Garth L.; Lynch, Sasha L.; Ahn, Yong-Mo; Ambler, Jon; Young, Katherine; Crowley, Brendan M.; Olsen, David B.; Warner, Digby F.; Barry, Clifton E.; Boshoff, Helena I.; Lamers, Meindert H.; Mizrahi, Valerie

doi:10.1021/acsinfecdis.1c00643

Cited by 14 publications

(11 citation statements)

References 80 publications

(175 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compared to viral and bacteriophage replicative DNA polymerases, mechanistic studies of cellular replicative polymerases have lagged behind. Among the cellular replicative polymerases, the bacterial C-family polymerases have been particularly challenging to study, largely because of weak binding to DNA and the unique catalytic cycle of the enzyme, but the available structures of several essential C-family polymerases (7,8,(31)(32)(33)(34)(35)(36) and our transientstate kinetic characterization of S. aureus PolC (16,17,24) provide the framework needed for a mechanistic structure-function approach to developing antibiotics targeting bacterial DNA replication.…”

Section: Discussionmentioning

confidence: 99%

Pre-Steady-State Kinetic Characterization of an Antibiotic-Resistant Mutation of Staphylococcus aureus DNA Polymerase PolC

Nelson-Rigg

Fagan

Jaremko

et al. 2022

Preprint

View full text Add to dashboard Cite

The emergence and spread of antibiotic resistance in bacterial pathogens are serious and ongoing threats to public health. Since chromosome replication is essential to cell growth and pathogenesis, the essential DNA polymerases in bacteria have long been targets of antimicrobial development, although none have yet advanced to the market. Here we use transient-state kinetic methods to characterize the inhibition of the PolC replicative DNA polymerase from Staphylococcus aureus by ME-EMAU, a member of the 6-anilinouracil compounds that specifically target PolC enzymes, which are found in low-GC content Gram- positive bacteria. We find that ME-EMAU binds to S. aureus PolC with a dissociation constant of 14 nM, more than 200-fold tighter than the previously reported inhibition constant, which was determined using steady-state kinetic methods. This tight binding is driven by a very slow off rate, 0.006 s-1. We also characterized the kinetics of nucleotide incorporation by PolC containing a mutation of phenylalanine 1261 to leucine (F1261L). The F1261L mutation decreases ME-EMAU binding affinity by at least 3500-fold, but also decreases the maximal rate of nucleotide incorporation by 11.5-fold. This suggests that bacteria acquiring this mutation would be likely to replicate slowly and be unable to out-compete wild-type strains in the absence of inhibitor, reducing the likelihood of the resistant bacteria propagating and spreading resistance.

show abstract

Section: Discussionmentioning

confidence: 99%

Pre-Steady-State Kinetic Characterization of an Antibiotic-Resistant Mutation of Staphylococcus aureus DNA Polymerase PolC

Nelson-Rigg

Fagan

Jaremko

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…As deeply reviewed by Reiche et al (2017), the DNA replication machinery is encoded by essential mycobacterial genes. Thus, other replication inhibitors are being explored including compounds targeting topoisomerase I (Sandhaus et al, 2016;Ekins et al, 2017), DNA polymerase complex (by Nargenicin, targeting DnaE1) (Chengalroyen et al, 2022) or Griselymicin (DnaN) as described above. Whether or not these molecules targeting machinery other than the fluoroquinolone targeti.e.…”

Section: How Can We Rescue Compromised Validated Targetsmentioning

confidence: 99%

“Upcycling” known molecules and targets for drug-resistant TB

Roubert

Fontaine²,

Upton³

2022

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Despite reinvigorated efforts in Tuberculosis (TB) drug discovery over the past 20 years, relatively few new drugs and candidates have emerged with clear utility against drug resistant TB. Over the same period, significant technological advances and learnings around target value have taken place. This has offered opportunities to re-assess the potential for optimization of previously discovered chemical matter against Mycobacterium tuberculosis (M.tb) and for reconsideration of clinically validated targets encumbered by drug resistance. A re-assessment of discarded compounds and programs from the “golden age of antibiotics” has yielded new scaffolds and targets against TB and uncovered classes, for example beta-lactams, with previously unappreciated utility for TB. Leveraging validated classes and targets has also met with success: booster technologies and efforts to thwart efflux have improved the potential of ethionamide and spectinomycin classes. Multiple programs to rescue high value targets while avoiding cross-resistance are making progress. These attempts to make the most of known classes, drugs and targets complement efforts to discover new chemical matter against novel targets, enhancing the chances of success of discovering effective novel regimens against drug-resistant TB.

show abstract

“…During host infection, the increase of DNA gene expression directly contributes to mutation leads to DNA damage induced by immunemediated reactive oxygen and nitrogen intermediates. Especially, ATP genes may play an essential role in the emergence of mutants that are better suited to surviving during infection and drug resistance evolution in this organism [29]. The bacteria tend to reduce cellular ATP consumption while increasing the capacity of ATP-generating pathways, which adds to bacterial survival in the face of antibiotic stress.…”

Section: Comprehensive Analysis Of Mirna With Transcription Factormentioning

confidence: 99%

An in silico analysis: Deciphering the role of differential expressed genes in tuberculosis in response to bedaquiline antibiotic drug

Santhiya¹,

Anusuya²,

Madhan³

et al. 2022

J App Biol biotech

View full text Add to dashboard Cite

Tuberculosis (TB) caused by the bacteria Mycobacterium TB (MTB) is one of the most devastating respiratory infections. The drug Bedaquiline, which is used to treat tuberculosis, was developed to improve treatment outcomes while reducing the toxicity associated with injectable medicines. Clinical prescriptions for the medication have increased in recent years, but its therapeutic efficacy has decreased. Drug resistance arises when MTB become resistant to the medications used to treat TB, and these treatments no longer suppress the gene activity. In the present study, an integrated bioinformatics analysis was performed to explore potential crucial genes, key pathways and interaction of miRNA with the transcription factor associated with the differentially expressed gene in TB in response to a multidrug resistant TB drug named bedaquiline. We have curated the dataset GSE43764 and carried out the functional and pathway enrichment analysis using various bioinformatics tool. The genes encoding dnaA, dnaB, dnaE1, atpE, atpF, and atpB were identified as hub genes. MiR-574-3p, MiR-4787-5p, MiR-601, and MiR-1234 have been found as possible target microRNA. The potential transcription factors pertaining to the Bedaquiline and TB were also found. The results indicate that the discovered key differentially expressed gene has a promising future and can be employed as biomarkers.

show abstract

DNA-Dependent Binding of Nargenicin to DnaE1 Inhibits Replication in Mycobacterium tuberculosis

Cited by 14 publications

References 80 publications

Pre-Steady-State Kinetic Characterization of an Antibiotic-Resistant Mutation of Staphylococcus aureus DNA Polymerase PolC

Pre-Steady-State Kinetic Characterization of an Antibiotic-Resistant Mutation of Staphylococcus aureus DNA Polymerase PolC

“Upcycling” known molecules and targets for drug-resistant TB

An in silico analysis: Deciphering the role of differential expressed genes in tuberculosis in response to bedaquiline antibiotic drug

Contact Info

Product

Resources

About