Heterogeneous fitness landscape cues, pknG low expression, and phthiocerol dimycocerosate low production of Mycobacterium tuberculosis ATCC25618 rpoB S450L in enriched broth

Édgar Rodríguez-Beltrán; Gerson-Dirceu López; Juan Manuel Anzola; Juan Germán Rodríguez-Castillo; Chiara Carazzone; Martha I Murcia

doi:10.1016/j.tube.2021.102156

Heterogeneous fitness landscape cues, pknG low expression, and phthiocerol dimycocerosate low production of Mycobacterium tuberculosis ATCC25618 rpoB S450L in enriched broth

Tuberculosis (Edinb). 2022 Jan:132:102156. doi: 10.1016/j.tube.2021.102156. Epub 2021 Dec 3.

Authors

Édgar Rodríguez-Beltrán¹, Gerson-Dirceu López², Juan Manuel Anzola³, Juan Germán Rodríguez-Castillo¹, Chiara Carazzone², Martha I Murcia⁴

Affiliations

¹ MicobacUN Group, Microbiology Department, The National University of Colombia (NUC) School of Medicine, AV CR 30 45-03, Bogotá, D.C, 111321, Colombia.
² Laboratory of Advanced Analytical Techniques in Natural Products (LATNAP), Chemistry Department, Universidad de los Andes, CR 1 18A-12, Bogotá, D.C, 111711, Colombia.
³ Corpogen, CR 4 20-41, Bogotá, D.C, 110311, Colombia; Universidad Central, CR 5 21-38, Bogotá, D.C, 110311, Colombia.
⁴ MicobacUN Group, Microbiology Department, The National University of Colombia (NUC) School of Medicine, AV CR 30 45-03, Bogotá, D.C, 111321, Colombia. Electronic address: mimurciaa@unal.edu.co.

PMID: 34891037
DOI: 10.1016/j.tube.2021.102156

Abstract

Multidrug-resistant tuberculosis (isoniazid/rifampin[RIF]-resistant TB) ravages developing countries. Fitness is critical in clinical outcomes. Previous studies on RIF-resistant TB (RR-TB) showed competitive fitness gains and losses, with rpoB-S450L as the most isolated/fit mutation. This study measured virulence/resistance genes, phthiocerol dimycocerosate (PDIM) levels and their relationship with rpoB S450L ATCC25618 RR-TB strain fitness. After obtaining 10 different RR-TB GenoType MTBDRplus 2.0-genotyped isolates (with nontyped, S441, H445 and S450 positions), only one S450L isolate (R9, rpoB-S450L ATCC 25618, RR 1 μg/mL) was observed, with H445Y being the most common. A competitive fitness in vitro assay with wild-type (wt) ATCC 25618: R9 1:1 in 50 mL Middlebrook 7H9/OADC was performed, and generation time (G) in vitro and relative fitness were obtained. mRNA and PDIM were extracted on log and stationary phases. Fitness decreased in rpoB S450L and H445Y strains, with heterogeneous fitness cues in three biological replicas of rpoB-S450L: one high and two low fitness replicas. S450L strain had significant pknG increase. Compared with S450L, wt-rpoB showed increased polyketide synthase ppsA expression and high PDIM peak measured by HPLC-MS in log phase compared to S450L. This contrasts with previously increased PDIM in other RR-TB isolates.

Keywords: Fitness; PDIM; Rifampicin; Tuberculosis; Virulence; rpoB.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antitubercular Agents / metabolism
Antitubercular Agents / therapeutic use
Bacterial Proteins / metabolism*
Humans
Lipids / biosynthesis*
Mycobacterium tuberculosis / genetics
Protein Serine-Threonine Kinases / metabolism*
Rifampin / metabolism
Rifampin / therapeutic use
Tuberculosis, Multidrug-Resistant / genetics*

Substances

Antitubercular Agents
Bacterial Proteins
Lipids
phthiocerol dimycocerosate
Protein Serine-Threonine Kinases
protein kinase G, Mycobacterium tuberculosis
Rifampin