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Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo
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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9045647/ https://www.ncbi.nlm.nih.gov/pubmed/35427399 http://dx.doi.org/10.1371/journal.ppat.1010475 |
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author | Tripathi, Ashutosh Anand, Kushi Das, Mayashree O’Niel, Ruchika Annie P. S, Sabarinath Thakur, Chandrani R. L., Raghunatha Reddy Rajmani, Raju S. Chandra, Nagasuma Laxman, Sunil Singh, Amit |
author_facet | Tripathi, Ashutosh Anand, Kushi Das, Mayashree O’Niel, Ruchika Annie P. S, Sabarinath Thakur, Chandrani R. L., Raghunatha Reddy Rajmani, Raju S. Chandra, Nagasuma Laxman, Sunil Singh, Amit |
author_sort | Tripathi, Ashutosh |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9045647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-90456472022-04-28 Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo Tripathi, Ashutosh Anand, Kushi Das, Mayashree O’Niel, Ruchika Annie P. S, Sabarinath Thakur, Chandrani R. L., Raghunatha Reddy Rajmani, Raju S. Chandra, Nagasuma Laxman, Sunil Singh, Amit PLoS Pathog Research Article 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. Public Library of Science 2022-04-15 /pmc/articles/PMC9045647/ /pubmed/35427399 http://dx.doi.org/10.1371/journal.ppat.1010475 Text en © 2022 Tripathi et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Tripathi, Ashutosh Anand, Kushi Das, Mayashree O’Niel, Ruchika Annie P. S, Sabarinath Thakur, Chandrani R. L., Raghunatha Reddy Rajmani, Raju S. Chandra, Nagasuma Laxman, Sunil Singh, Amit Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title | Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title_full | Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title_fullStr | Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title_full_unstemmed | Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title_short | Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo |
title_sort | mycobacterium tuberculosis requires suft for fe-s cluster maturation, metabolism, and survival in vivo |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9045647/ https://www.ncbi.nlm.nih.gov/pubmed/35427399 http://dx.doi.org/10.1371/journal.ppat.1010475 |
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