The impact of genotype on the phenotype of Mycobacterium tuberculosis ΔsufR mutants

Danicke Willemse; Lucinda Baatjies; Anzaan Dippenaar; Robin M Warren; Monique J Williams

doi:10.1016/j.tube.2023.102360

The impact of genotype on the phenotype of Mycobacterium tuberculosis ΔsufR mutants

Tuberculosis (Edinb). 2023 Jul:141:102360. doi: 10.1016/j.tube.2023.102360. Epub 2023 Jun 2.

Authors

Danicke Willemse¹, Lucinda Baatjies¹, Anzaan Dippenaar², Robin M Warren¹, Monique J Williams³

Affiliations

¹ Department of Science and Innovation (DSI)-National Research Foundation (NRF) Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa.
² Department of Science and Innovation (DSI)-National Research Foundation (NRF) Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa; Tuberculosis Omics Research Consortium, Family Medicine and Population Health, Institute of Global Health, Faculty of Medicine and Health Sciences, University of Antwerp, Antwerp, Belgium.
³ Department of Science and Innovation (DSI)-National Research Foundation (NRF) Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa; Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa. Electronic address: monique.williams@uct.ac.za.

PMID: 37295353
DOI: 10.1016/j.tube.2023.102360

Abstract

Iron-sulphur (FeS) cluster biogenesis is a tightly regulated process in vivo. In Mycobacterium tuberculosis (Mtb), SufR functions as a transcriptional repressor of the operon encoding the primary FeS cluster biogenesis system. Previously, three independently isolated mutants (ΔRv1460stop_1.19, ΔRv1460stop _5.19 and ΔRv1460stop _5.20) harbouring the same deletion in sufR, displayed different growth kinetics in OADC supplemented 7H9 media. To investigate this discrepancy, we performed whole genome sequencing of the 3 mutants and the wild-type progenitor. Single nucleotide polymorphisms (SNPs) were identified in 3 genes in the ΔRv1460stop_1.19 mutant and one gene in the ΔRv1460stop_5.20 mutant. Phenotyping of the ΔRv1460stop_5.19 mutant, which had no additional SNPs, revealed increased susceptibility to clofazimine, DMNQ and menadione, while uptake and survival in THP-1 cells were not significantly different from the wild-type strain. Given that these results differ from those reported for other sufR deletion mutants (ΔSufR_MTB and MtbΔSufR), they suggest that the position of the sufR deletion and the genotype of the progenitor strain impact the resulting phenotype.

Keywords: Iron-sulphur cluster; Mycobacterium tuberculosis; Regulation; SufR; rv1460.

Publication types

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

MeSH terms

Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Genotype
Iron-Sulfur Proteins* / genetics
Mycobacterium tuberculosis*
Phenotype

Substances

Iron-Sulfur Proteins
Bacterial Proteins