Evaluating the Sensitivity of Mycobacterium tuberculosis to Biotin Deprivation Using Regulated Gene Expression

• DOI: 10.1371/journal.ppat.1002264
• Publication Date: 2011-09-29T17:37:31Z
• Authors: Sae Woong Park; Marcus Klotzsche; Daniel J. Wilson; Helena I. Boshoff; Hyungjin Eoh; Ujjini Manjunatha; Antje Blumenthal; Kyu Rhee; Clifton E. Barry; Courtney C. Aldrich; Sabine Ehrt; Dirk Schnappinger
• Keywords: 0301 basic medicine; protein synthesis; Drug targets; Biotin synthetic pathway; 2405 Parasitology; bacterial growth; bacterial protein; gene silencing; Mice; Drug Delivery Systems; sensitivity analysis; biotin; drug delivery system; animal; genetics; biotinylation; Biology (General); bacteria; Bacteria (microorganisms); quantitative analysis; Mus; 2404 Microbiology; article; methodology; gene expression regulation; biotin derivative; unclassified drug; enzyme activity; 3. Good health; tuberculosis; wild type; signal transduction; Research Article; 570; in vitro study; QH301-705.5; enzymology; animal experiment; 610; Biotin; animal tissue; in vivo study; 03 medical and health sciences; BioA; 1311 Genetics; Bacterial Proteins; 1312 Molecular Biology; Animals; Tuberculosis; anhydrotetracycline; controlled study; BioA protein; 7,8 diaminopelargonic acid synthase; mouse; Transaminases; Bacterial growth; 2403 Immunology; nonhuman; doxycycline; aminotransferase; animal model; disease model; bacterial viability; Mycobacterium tuberculosis; RC581-607; Disease Models, Animal; Chronic Disease; Mutation; 2406 Virology; Murinae; biosynthesis; mutation; Immunologic diseases. Allergy; chronic disease; metabolism

In the search for new drug targets, we evaluated biotin synthetic pathway of Mycobacterium tuberculosis (Mtb) and constructed an Mtb mutant lacking biosynthetic enzyme 7,8-diaminopelargonic acid synthase, BioA. biotin-free media, ΔbioA did not produce wild-type levels biotinylated proteins, therefore grow lost viability. was also unable to establish infection in mice. Conditionally-regulated knockdown strains similarly exhibited impaired bacterial growth viability vitro mice, irrespective timing transcriptional silencing. Biochemical studies further showed that BioA activity has be reduced by approximately 99% prevent growth. These thus de novo synthesis is essential maintain a chronic murine model TB. Moreover, these provide experimental strategy systematically rank vivo value potential targets other pathogens.