title: Menaquinone synthesis is critical for maintaining mycobacterial viability during exponential growth and recovery from non-replicating persistence.
abstract: Understanding the basis of bacterial persistence in latent infections is critical for eradication of tuberculosis. Analysis of Mycobacterium tuberculosis mRNA expression in an in vitro model of non-replicating persistence indicated that the bacilli require electron transport chain components and ATP synthesis for survival. Additionally, low microM concentrations of aminoalkoxydiphenylmethane derivatives inhibited both the aerobic growth and survival of non-replicating, persistent M. tuberculosis. Metabolic labelling studies and quantification of cellular menaquinone levels suggested that menaquinone synthesis, and consequently electron transport, is the target of the aminoalkoxydiphenylmethane derivatives. This hypothesis is strongly supported by the observations that treatment with these compounds inhibits oxygen consumption and that supplementation of growth medium with exogenous menaquinone rescued both growth and oxygen consumption of treated bacilli. In vitro assays indicate that the aminoalkoxydiphenylmethane derivatives specifically inhibit MenA, an enzyme involved in the synthesis of menaquinone. Thus, the results provide insight into the physiology of mycobacterial persistence and a basis for the development of novel drugs that enhance eradication of persistent bacilli and latent tuberculosis.
authors.affiliation: Colorado State University, Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, 1682 Campus Delivery, Fort Collins, CO 80523, USA.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Dhiman
authors.forename: Sebabrata
authors.initials: S
authors.lastname: Mahapatra
authors.forename: Richard A
authors.initials: RA
authors.lastname: Slayden
authors.forename: Melissa E
authors.initials: ME
authors.lastname: Boyne
authors.forename: Anne
authors.initials: A
authors.lastname: Lenaerts
authors.forename: Jerald C
authors.initials: JC
authors.lastname: Hinshaw
authors.forename: Shiva K
authors.initials: SK
authors.lastname: Angala
authors.forename: Delphi
authors.initials: D
authors.lastname: Chatterjee
authors.forename: Kallolmay
authors.initials: K
authors.lastname: Biswas
authors.forename: Prabagaran
authors.initials: P
authors.lastname: Narayanasamy
authors.forename: Michio
authors.initials: M
authors.lastname: Kurosu
authors.forename: Dean C
authors.initials: DC
authors.lastname: Crick
mesh_terms: D000255:Adenosine Triphosphate / Q000096:biosynthesis; D045222:Electron Transport Chain Complex Proteins / Q000096:biosynthesis; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D050296:Microbial Viability; D009169:Mycobacterium tuberculosis / Q000254:growth & development* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010101:Oxygen Consumption; D012329:RNA, Bacterial / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D024482:Vitamin K 2 / Q000378:metabolism*
chemical_list: D045222:Electron Transport Chain Complex Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D024482:Vitamin K 2; D000255:Adenosine Triphosphate