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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2022-04-01
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| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1010475 |
| _version_ | 1818478688819216384 |
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| author | Ashutosh Tripathi Kushi Anand Mayashree Das Ruchika Annie O'Niel Sabarinath P S Chandrani Thakur Raghunatha Reddy R L Raju S Rajmani Nagasuma Chandra Sunil Laxman Amit Singh |
| author_facet | Ashutosh Tripathi Kushi Anand Mayashree Das Ruchika Annie O'Niel Sabarinath P S Chandrani Thakur Raghunatha Reddy R L Raju S Rajmani Nagasuma Chandra Sunil Laxman Amit Singh |
| author_sort | Ashutosh Tripathi |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-10T09:50:52Z |
| format | Article |
| id | doaj.art-dbdb527b1f54415395c8109b4e42af7c |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-10T09:50:52Z |
| publishDate | 2022-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-dbdb527b1f54415395c8109b4e42af7c2022-12-22T01:53:40ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742022-04-01184e101047510.1371/journal.ppat.1010475Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo.Ashutosh TripathiKushi AnandMayashree DasRuchika Annie O'NielSabarinath P SChandrani ThakurRaghunatha Reddy R LRaju S RajmaniNagasuma ChandraSunil LaxmanAmit SinghIron-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.https://doi.org/10.1371/journal.ppat.1010475 |
| spellingShingle | Ashutosh Tripathi Kushi Anand Mayashree Das Ruchika Annie O'Niel Sabarinath P S Chandrani Thakur Raghunatha Reddy R L Raju S Rajmani Nagasuma Chandra Sunil Laxman Amit Singh Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo. PLoS Pathogens |
| 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 |
| url | https://doi.org/10.1371/journal.ppat.1010475 |
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