Boshoff, Helena Ingrid and Tripathi, Ashutosh and Anand, Kushi and Das, Mayashree and O’Niel, Ruchika Annie and PS, Sabarinath and Thakur, Chandrani and RL, Raghunatha Reddy and Rajmani, Raju S and Chandra, Nagasuma and Laxman, Sunil and Singh, Amit (2022) Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo. In: PLOS Pathogens, 18 (4). ISSN 1553-7374
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Abstract
Iron-sulfur (Fe-S) cluster proteins carry out essential cellular functions in diverse organisms, including the human pathogen Mycobacterium tuberculosis (Mtb). The mechanisms underlying Fe-S cluster biogenesis are poorly defined in Mtb. Here, we show that Mtb SufT (Rv1466), a DUF59 domain-containing essential protein, is required for the Fe-S cluster maturation. Mtb SufT homodimerizes and interacts with Fe-S cluster biogenesis proteins; SufS and SufU. SufT also interacts with the 4Fe-4S cluster containing proteins; aconitase and SufR. Importantly, a hyperactive cysteine in the DUF59 domain mediates interaction of SufT with SufS, SufU, aconitase, and SufR. We efficiently repressed the expression of SufT to generate a SufT knock-down strain in Mtb (SufT-KD) using CRISPR interference. Depleting SufT reduces aconitase’s enzymatic activity under standard growth conditions and in response to oxidative stress and iron limitation. The SufT-KD strain exhibited defective growth and an altered pool of tricarboxylic acid cycle intermediates, amino acids, and sulfur metabolites. Using Seahorse Extracellular Flux analyzer, we demonstrated that SufT depletion diminishes glycolytic rate and oxidative phosphorylation in Mtb. The SufT-KD strain showed defective survival upon exposure to oxidative stress and nitric oxide. Lastly, SufT depletion reduced the survival of Mtb in macrophages and attenuated the ability of Mtb to persist in mice. Altogether, SufT assists in Fe-S cluster maturation and couples this process to bioenergetics of Mtb for survival under low and high demand for Fe-S clusters. © 2022 Tripathi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
| Item Type: | Journal Article |
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| Publication: | PLOS Pathogens |
| Publisher: | Public Library of Science |
| Additional Information: | The copyright of this article belongs to the Authors. |
| Department/Centre: | Division of Biological Sciences > Biochemistry |
| Date Deposited: | 23 May 2022 10:31 |
| Last Modified: | 23 May 2022 10:31 |
| URI: | https://eprints.iisc.ac.in/id/eprint/72364 |
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