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Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence

Virulence effectors secreted by Mycobacterium tuberculosis (Mtb) help subvert host immune mechanisms and, therefore, are critical for establishing infection and pathogenesis. However, knowledge in terms of signaling mechanisms that modulate the secretion of virulence factors is sparse. We performed...

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Autores principales: Malakar, Basanti, Chauhan, Komal, Sanyal, Priyadarshini, Naz, Saba, Kalam, Haroon, Vivek-Ananth, R. P., Singh, Lakshya Veer, Samal, Areejit, Kumar, Dhiraj, Nandicoori, Vinay Kumar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10653824/
https://www.ncbi.nlm.nih.gov/pubmed/37791794
http://dx.doi.org/10.1128/mbio.01232-23
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author Malakar, Basanti
Chauhan, Komal
Sanyal, Priyadarshini
Naz, Saba
Kalam, Haroon
Vivek-Ananth, R. P.
Singh, Lakshya Veer
Samal, Areejit
Kumar, Dhiraj
Nandicoori, Vinay Kumar
author_facet Malakar, Basanti
Chauhan, Komal
Sanyal, Priyadarshini
Naz, Saba
Kalam, Haroon
Vivek-Ananth, R. P.
Singh, Lakshya Veer
Samal, Areejit
Kumar, Dhiraj
Nandicoori, Vinay Kumar
author_sort Malakar, Basanti
collection PubMed
description Virulence effectors secreted by Mycobacterium tuberculosis (Mtb) help subvert host immune mechanisms and, therefore, are critical for establishing infection and pathogenesis. However, knowledge in terms of signaling mechanisms that modulate the secretion of virulence factors is sparse. We performed high-throughput secretome, phosphoproteome, and phospho-secretome analysis of Mtb. We combined the analysis with empirical validations to show regulation of mycobacterial secretion through protein phosphorylation. System level protein-protein interaction network analysis superimposed with the secretome, phosphoproteome, and phospho-secretome profile revealed an intricate relationship between phosphorylation and secretion. At the core of the network was a key virulence factor CFP10. We identified PknA to be the kinase responsible for phosphorylating CFP10. Using genetic tools, we show that phosphomimetic mutation of CFP10 negatively regulates the secretion of virulence mediator ESAT6. Significantly, the dynamics of CFP10 phosphorylation strongly influenced bacterial virulence and survival within macrophages and mice. Together, the results show that the dynamic phosphorylation status of the secretory protein CFP10 regulates the secretion of virulence factors and impacts virulence. IMPORTANCE: Secreted virulence factors play a critical role in bacterial pathogenesis. Virulence effectors not only help bacteria to overcome the host immune system but also aid in establishing infection. Mtb, which causes tuberculosis in humans, encodes various virulence effectors. Triggers that modulate the secretion of virulence effectors in Mtb are yet to be fully understood. To gain mechanistic insight into the secretion of virulence effectors, we performed high-throughput proteomic studies. With the help of system-level protein-protein interaction network analysis and empirical validations, we unravelled a link between phosphorylation and secretion. Taking the example of the well-known virulence factor of CFP10, we show that the dynamics of CFP10 phosphorylation strongly influenced bacterial virulence and survival ex vivo and in vivo. This study presents the role of phosphorylation in modulating the secretion of virulence factors.
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spelling pubmed-106538242023-10-04 Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence Malakar, Basanti Chauhan, Komal Sanyal, Priyadarshini Naz, Saba Kalam, Haroon Vivek-Ananth, R. P. Singh, Lakshya Veer Samal, Areejit Kumar, Dhiraj Nandicoori, Vinay Kumar mBio Research Article Virulence effectors secreted by Mycobacterium tuberculosis (Mtb) help subvert host immune mechanisms and, therefore, are critical for establishing infection and pathogenesis. However, knowledge in terms of signaling mechanisms that modulate the secretion of virulence factors is sparse. We performed high-throughput secretome, phosphoproteome, and phospho-secretome analysis of Mtb. We combined the analysis with empirical validations to show regulation of mycobacterial secretion through protein phosphorylation. System level protein-protein interaction network analysis superimposed with the secretome, phosphoproteome, and phospho-secretome profile revealed an intricate relationship between phosphorylation and secretion. At the core of the network was a key virulence factor CFP10. We identified PknA to be the kinase responsible for phosphorylating CFP10. Using genetic tools, we show that phosphomimetic mutation of CFP10 negatively regulates the secretion of virulence mediator ESAT6. Significantly, the dynamics of CFP10 phosphorylation strongly influenced bacterial virulence and survival within macrophages and mice. Together, the results show that the dynamic phosphorylation status of the secretory protein CFP10 regulates the secretion of virulence factors and impacts virulence. IMPORTANCE: Secreted virulence factors play a critical role in bacterial pathogenesis. Virulence effectors not only help bacteria to overcome the host immune system but also aid in establishing infection. Mtb, which causes tuberculosis in humans, encodes various virulence effectors. Triggers that modulate the secretion of virulence effectors in Mtb are yet to be fully understood. To gain mechanistic insight into the secretion of virulence effectors, we performed high-throughput proteomic studies. With the help of system-level protein-protein interaction network analysis and empirical validations, we unravelled a link between phosphorylation and secretion. Taking the example of the well-known virulence factor of CFP10, we show that the dynamics of CFP10 phosphorylation strongly influenced bacterial virulence and survival ex vivo and in vivo. This study presents the role of phosphorylation in modulating the secretion of virulence factors. American Society for Microbiology 2023-10-04 /pmc/articles/PMC10653824/ /pubmed/37791794 http://dx.doi.org/10.1128/mbio.01232-23 Text en Copyright © 2023 Malakar et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Malakar, Basanti
Chauhan, Komal
Sanyal, Priyadarshini
Naz, Saba
Kalam, Haroon
Vivek-Ananth, R. P.
Singh, Lakshya Veer
Samal, Areejit
Kumar, Dhiraj
Nandicoori, Vinay Kumar
Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title_full Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title_fullStr Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title_full_unstemmed Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title_short Phosphorylation of CFP10 modulates Mycobacterium tuberculosis virulence
title_sort phosphorylation of cfp10 modulates mycobacterium tuberculosis virulence
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10653824/
https://www.ncbi.nlm.nih.gov/pubmed/37791794
http://dx.doi.org/10.1128/mbio.01232-23
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