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A high-throughput whole cell screen to identify inhibitors of Mycobacterium tuberculosis

for a variety of reasons including lack of permeation, efflux, and poor target vulnerability or engagement [16]. Competing interests: Tanya Parish serves on the Editorial Board of PLOS ONE. This does ... external funding received for this study. Author Contributions Conceptualization: Juliane Ollinger, Anuradha Kumar, David M. Roberts, Mai A. Bailey, Tanya Parish. Data curation: Anuradha Kumar, David M

Codon-optimized DsRed fluorescent protein for use in Mycobacterium tuberculosis

ObjectiveWe have previously codon-optimized a number of red fluorescent proteins for use in Mycobacterium tuberculosis (mCherry, tdTomato, Turbo-635). We aimed to expand this repertoire to include DsRed, another widely used and flexible red fluorescent protein.ResultsWe generated expression constructs with a full length DsRed under the control of one of three strong, constitutive...

A class of hydrazones are active against non-replicating Mycobacterium tuberculosis

, WA , United States of America 2 Competing interests: Tanya Parish serves on the Editorial Board of PLOS ONE. This does not alter There is an urgent need for the development of shorter, simpler and ... . Author Contributions Conceptualization: Shilah A. Bonnett, Devon Dennison, Tanya Parish. Data curation: Shilah A. Bonnett, Tanya Parish. Formal analysis: Shilah A. Bonnett, Devon Dennison, Megan Files

High content, high-throughput screening for small molecule inducers of NF-κB translocation

Parish. Data curation: Samuel Njikan, Yulia Ovechkina, Divya Awasthi, Tanya Parish. Formal analysis: Samuel Njikan, Yulia Ovechkina, Divya Awasthi, Tanya Parish. Funding acquisition: Tanya Parish ... . Investigation: Samuel Njikan, Alyssa J. Manning. Methodology: Samuel Njikan, Alyssa J. Manning, Yulia Ovechkina, Divya Awasthi. Project administration: Tanya Parish. Resources: Tanya Parish. Supervision: Tanya

Nitazoxanide is active against Mycobacterium leprae

. Taken together, this information and our data suggest that NTZ could be considered as a supplement to the drug arsenal for the treatment of leprosy. Author Contributions Tanya Parish. Data curation ... : Tanya Parish. Conceptualization: Mai Ann Bailey, Malcolm S. Duthie, Thomas P. Gillis, Ramanuj Lahiri, Formal analysis: Malcolm S. Duthie, Thomas P. Gillis, Ramanuj Lahiri, Tanya Parish. Funding

Deletion of cyp125 Confers Increased Sensitivity to Azoles in Mycobacterium tuberculosis

Mycobacterium tuberculosis is able to utilize cholesterol as a carbon source, and this ability is linked to its virulence in macrophages and in the mouse model of infection. The M. tuberculosis cytochrome P450 Cyp125 plays a key role in cholesterol metabolism being involved in the first steps of its degradation. Cyp125 is a cholesterol hydroxylase which is essential for...

LytB1 and LytB2 of Mycobacterium tuberculosis Are Not Genetically Redundant

Mycobacterium tuberculosis synthesises isoprenoid precursors via the MEP/DOXP pathway and at least five enzymes in the pathway (Dxs1, Dxr/IspC, IspD, IspF, and GcpE/IspG) are required for growth in vitro. We investigated the role of LytB (IspH) in M. tuberculosis; M. tuberculosis is unusual in that it has two homologs–LytB1 and LytB2. We were unable to delete the lytB2 gene...

Mycobacterium tuberculosis DosR is Required for Activity of the PmbtB and PmbtI Promoters under Hypoxia

Mycobacterium tuberculosis has the ability to survive for extended periods of time under conditions of low oxygen, low pH, low iron and low nutrients. The mycobactins (M. tuberculosis siderophores) play a key role in scavenging iron from the environment and are induced in response to low iron in an IdeR-regulated manner. We demonstrate that the promoters of two mycobactin gene...

Identification of the translational start site of codon-optimized mCherry in Mycobacterium tuberculosis

Tanya Parish 0 1 0 Infectious Disease Research Institute , Seattle, Washington , USA 1 Queen Mary University of London, Barts & The London School of Medicine and Dentistry, Centre for Immunology and

Identification of the likely translational start of Mycobacterium tuberculosis GyrB

Background Bacterial DNA gyrase is a validated target for antibacterial chemotherapy. It consists of two subunits, GyrA and GyrB, which form an A2B2 complex in the active enzyme. Sequence alignment of Mycobacterium tuberculosis GyrB with other bacterial GyrBs predicts the presence of 40 potential additional amino acids at the GyrB N-terminus. There are discrepancies between the M...

The Promoter of Rv0560c Is Induced by Salicylate and Structurally-Related Compounds in Mycobacterium tuberculosis

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a major global health threat. During infection, bacteria are believed to encounter adverse conditions such as iron depletion. Mycobacteria synthesize iron-sequestering mycobactins, which are essential for survival in the host, via the intermediate salicylate. Salicylate is a ubiquitous compound which is...

Synthesis and Evaluation of the 2-Aminothiazoles as Anti-Tubercular Agents

The 2-aminothiazole series has anti-bacterial activity against the important global pathogen Mycobacterium tuberculosis. We explored the nature of the activity by designing and synthesizing a large number of analogs and testing these for activity against M. tuberculosis, as well as eukaryotic cells. We determined that the C-2 position of the thiazole can accommodate a range of...

Mycobacterium tuberculosis ClpP Proteases Are Co-transcribed but Exhibit Different Substrate Specificities

therefore have different, but overlapping roles in M. tuberculosis. - Competing Interests: Tanya Parish served on the Editorial Board of PLOS ONE. This does not alter the authors adherence to all the PLOS

Mycobacterium tuberculosis Responds to Chloride and pH as Synergistic Cues to the Immune Status of its Host Cell

The ability of Mycobacterium tuberculosis (Mtb) to thrive in its phagosomal niche is critical for its establishment of a chronic infection. This requires that Mtb senses and responds to intraphagosomal signals such as pH. We hypothesized that Mtb would respond to additional intraphagosomal factors that correlate with maturation. Here, we demonstrate that [Cl−] and pH correlate...

Control of CydB and GltA1 Expression by the SenX3 RegX3 Two Component Regulatory System of Mycobacterium tuberculosis

Two component regulatory systems are used widely by bacteria to coordinate changes in global gene expression profiles in response to environmental signals. The SenX3-RegX3 two component system of Mycobacterium tuberculosis has previously been shown to play a role in virulence and phosphate-responsive control of gene expression. We demonstrate that expression of SenX3-RegX3 is...

A Dual Read-Out Assay to Evaluate the Potency of Compounds Active against Mycobacterium tuberculosis

design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: Tanya Parish serves on the Editorial Board of PLOS ONE. This does not alter the authors ... Parish 0 Petros C. Karakousis, Johns Hopkins University School of Medicine, United States of America 0 1 Infectious Disease Research Institute, Seattle, Washington, United States of America, 2 Department

Dxr is essential in Mycobacterium tuberculosis and fosmidomycin resistance is due to a lack of uptake

Fosmidomycin is a phosphonic antibiotic which inhibits 1-deoxy-D-xylulose 5-phosphate reductoisomerase (Dxr), the first committed step of the non-mevalonate pathway of isoprenoid biosynthesis. In Mycobacterium tuberculosis Dxr is encoded by Rv2870c, and although the antibiotic has been shown to inhibit the recombinant enzyme 1, mycobacteria are intrinsically resistant to...

The dUTPase Enzyme Is Essential in Mycobacterium smegmatis

Thymidine biosynthesis is essential in all cells. Inhibitors of the enzymes involved in this pathway (e.g. methotrexate) are thus frequently used as cytostatics. Due to its pivotal role in mycobacterial thymidylate synthesis dUTPase, which hydrolyzes dUTP into the dTTP precursor dUMP, has been suggested as a target for new antitubercular agents. All mycobacterial genomes encode...

A High-Throughput Screen against Pantothenate Synthetase (PanC) Identifies 3-Biphenyl-4-Cyanopyrrole-2-Carboxylic Acids as a New Class of Inhibitor with Activity against Mycobacterium tuberculosis

The enzyme pantothenate synthetase, PanC, is an attractive drug target in Mycobacterium tuberculosis. It is essential for the in vitro growth of M. tuberculosis and for survival of the bacteria in the mouse model of infection. PanC is absent from mammals. We developed an enzyme-based assay to identify inhibitors of PanC, optimized it for high-throughput screening, and tested a...

The structure of Mycobacteria 2C-methyl-D-erythritol-2,4-cyclodiphosphate synthase, an essential enzyme, provides a platform for drug discovery

Background The prevalence of tuberculosis, the prolonged and expensive treatment that this disease requires and an increase in drug resistance indicate an urgent need for new treatments. The 1-deoxy-D-xylulose 5-phosphate pathway of isoprenoid precursor biosynthesis is an attractive chemotherapeutic target because it occurs in many pathogens, including Mycobacterium tuberculosis...