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Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions

Tuberculosis, a lung disease caused by Mycobacterium tuberculosis (Mtb), is one of the ten leading causes of death worldwide affecting mainly developing countries. Mtb can persist and survive inside infected cells through modulation of host antibacterial attack, i.e., by avoiding the maturation of p...

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Autores principales: Forrellad, Marina Andrea, Blanco, Federico Carlos, Marrero Diaz de Villegas, Rubén, Vázquez, Cristina Lourdes, Yaneff, Agustín, García, Elizabeth Andrea, Gutierrez, Maximiliano Gabriel, Durán, Rosario, Villarino, Andrea, Bigi, Fabiana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7642983/
https://www.ncbi.nlm.nih.gov/pubmed/33193164
http://dx.doi.org/10.3389/fmicb.2020.570794
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author Forrellad, Marina Andrea
Blanco, Federico Carlos
Marrero Diaz de Villegas, Rubén
Vázquez, Cristina Lourdes
Yaneff, Agustín
García, Elizabeth Andrea
Gutierrez, Maximiliano Gabriel
Durán, Rosario
Villarino, Andrea
Bigi, Fabiana
author_facet Forrellad, Marina Andrea
Blanco, Federico Carlos
Marrero Diaz de Villegas, Rubén
Vázquez, Cristina Lourdes
Yaneff, Agustín
García, Elizabeth Andrea
Gutierrez, Maximiliano Gabriel
Durán, Rosario
Villarino, Andrea
Bigi, Fabiana
author_sort Forrellad, Marina Andrea
collection PubMed
description Tuberculosis, a lung disease caused by Mycobacterium tuberculosis (Mtb), is one of the ten leading causes of death worldwide affecting mainly developing countries. Mtb can persist and survive inside infected cells through modulation of host antibacterial attack, i.e., by avoiding the maturation of phagosome containing mycobacteria to more acidic endosomal compartment. In addition, bacterial phosphatases play a central role in the interplay between host cells and Mtb. In this study, we characterized the Rv2577 of Mtb as a potential alkaline phosphatase/phosphodiesterase enzyme. By an in vitro kinetic assay, we demonstrated that purified Rv2577 expressed in Mycobacterium smegmatis displays both enzyme activities, as evidenced by using the artificial substrates p-NPP and bis-(p-NPP). In addition, a three-dimensional model of Rv2577 allowed us to define the catalytic amino acid residues of the active site, which were confirmed by site-directed mutagenesis and enzyme activity analysis, being characteristic of a member of the metallophosphatase superfamily. Finally, a mutation introduced in Rv2577 reduced the replication of Mtb in mouse organs and impaired the arrest of phagosomes containing mycobacteria in early endosomes; which indicates Rv2577 plays a role in Mtb virulence.
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spelling pubmed-76429832020-11-13 Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions Forrellad, Marina Andrea Blanco, Federico Carlos Marrero Diaz de Villegas, Rubén Vázquez, Cristina Lourdes Yaneff, Agustín García, Elizabeth Andrea Gutierrez, Maximiliano Gabriel Durán, Rosario Villarino, Andrea Bigi, Fabiana Front Microbiol Microbiology Tuberculosis, a lung disease caused by Mycobacterium tuberculosis (Mtb), is one of the ten leading causes of death worldwide affecting mainly developing countries. Mtb can persist and survive inside infected cells through modulation of host antibacterial attack, i.e., by avoiding the maturation of phagosome containing mycobacteria to more acidic endosomal compartment. In addition, bacterial phosphatases play a central role in the interplay between host cells and Mtb. In this study, we characterized the Rv2577 of Mtb as a potential alkaline phosphatase/phosphodiesterase enzyme. By an in vitro kinetic assay, we demonstrated that purified Rv2577 expressed in Mycobacterium smegmatis displays both enzyme activities, as evidenced by using the artificial substrates p-NPP and bis-(p-NPP). In addition, a three-dimensional model of Rv2577 allowed us to define the catalytic amino acid residues of the active site, which were confirmed by site-directed mutagenesis and enzyme activity analysis, being characteristic of a member of the metallophosphatase superfamily. Finally, a mutation introduced in Rv2577 reduced the replication of Mtb in mouse organs and impaired the arrest of phagosomes containing mycobacteria in early endosomes; which indicates Rv2577 plays a role in Mtb virulence. Frontiers Media S.A. 2020-10-22 /pmc/articles/PMC7642983/ /pubmed/33193164 http://dx.doi.org/10.3389/fmicb.2020.570794 Text en Copyright © 2020 Forrellad, Blanco, Marrero Diaz de Villegas, Vázquez, Yaneff, García, Gutierrez, Durán, Villarino and Bigi. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Forrellad, Marina Andrea
Blanco, Federico Carlos
Marrero Diaz de Villegas, Rubén
Vázquez, Cristina Lourdes
Yaneff, Agustín
García, Elizabeth Andrea
Gutierrez, Maximiliano Gabriel
Durán, Rosario
Villarino, Andrea
Bigi, Fabiana
Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title_full Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title_fullStr Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title_full_unstemmed Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title_short Rv2577 of Mycobacterium tuberculosis Is a Virulence Factor With Dual Phosphatase and Phosphodiesterase Functions
title_sort rv2577 of mycobacterium tuberculosis is a virulence factor with dual phosphatase and phosphodiesterase functions
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7642983/
https://www.ncbi.nlm.nih.gov/pubmed/33193164
http://dx.doi.org/10.3389/fmicb.2020.570794
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