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

PLoS Pathogens

Courtney C. Aldrich¹  Daniel J. Wilson¹  Kyu Rhee²  Hyungjin Eoh²  Antje Blumenthal³  Sae Woong Park³  Sabine Ehrt³  Marcus Klotzsche³  Dirk Schnappinger³  Ujjini Manjunatha⁴  Clifton E. Barry III⁵  Helena I. Boshoff⁵ 

[1] Center for Drug Design, University of Minnesota, Minneapolis, Minnesota, United States of America;Department of Medicine, Weill Cornell Medical College, New York, New York, United States of America;Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America;Novartis Institute for Tropical Diseases, Singapore;Tuberculosis Research Section, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, United States of America

关键词: Mycobacterium tuberculosis;    Biotin;    Mouse models;    Immunoblotting;    Animal models of infection;    Biosynthesis;    Drug discovery;    Tuberculosis;   

DOI  :  10.1371/journal.ppat.1002264

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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

【 授权许可】

CC BY