Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>

ABSTRACT Trypanosoma cruzi is the agent of Chagas disease, and the finding that this parasite possesses a mitochondrial calcium uniporter (TcMCU) with characteristics similar to that of mammalian mitochondria was fundamental for the discovery of the molecular nature of MCU in eukaryotes. We report h...

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Autores principales:	Miguel A. Chiurillo, Noelia Lander, Mayara S. Bertolini, Melissa Storey, Anibal E. Vercesi, Roberto Docampo
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	calcium signaling mitochondria Trypanosoma uniporter Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2c66e00f1b9244cf888d9eea4da90625
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spelling	oai:doaj.org-article:2c66e00f1b9244cf888d9eea4da906252021-11-15T15:51:29ZDifferent Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>10.1128/mBio.00574-172150-7511https://doaj.org/article/2c66e00f1b9244cf888d9eea4da906252017-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00574-17https://doaj.org/toc/2150-7511ABSTRACT Trypanosoma cruzi is the agent of Chagas disease, and the finding that this parasite possesses a mitochondrial calcium uniporter (TcMCU) with characteristics similar to that of mammalian mitochondria was fundamental for the discovery of the molecular nature of MCU in eukaryotes. We report here that ablation of TcMCU, or its paralog TcMCUb, by clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 led to a marked decrease in mitochondrial Ca2+ uptake without affecting the membrane potential of these cells, whereas overexpression of each gene caused a significant increase in the ability of mitochondria to accumulate Ca2+. While TcMCU-knockout (KO) epimastigotes were viable and able to differentiate into trypomastigotes, infect host cells, and replicate normally, ablation of TcMCUb resulted in epimastigotes having an important growth defect, lower rates of respiration and metacyclogenesis, more pronounced autophagy changes under starvation, and significantly reduced infectivity. Overexpression of TcMCUb, in contrast to what was proposed for its mammalian ortholog, did not result in a dominant negative effect on TcMCU. IMPORTANCE The finding of a mitochondrial calcium uniporter (MCU) in Trypanosoma cruzi was essential for the discovery of the molecular nature of this transporter in mammals. In this work, we used the CRISPR/Cas9 technique that we recently developed for T. cruzi to knock out two components of the uniporter: MCU, the pore subunit, and MCUb, which was proposed as a negative regulator of MCU in human cells. In contrast to what occurs in human cells, MCU is not essential, while MCUb is essential for growth, differentiation, and infectivity; has a bioenergetic role; and does not act as a dominant negative subunit of MCU.Miguel A. ChiurilloNoelia LanderMayara S. BertoliniMelissa StoreyAnibal E. VercesiRoberto DocampoAmerican Society for Microbiologyarticlecalcium signalingmitochondriaTrypanosomauniporterMicrobiologyQR1-502ENmBio, Vol 8, Iss 3 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	calcium signaling mitochondria Trypanosoma uniporter Microbiology QR1-502
spellingShingle	calcium signaling mitochondria Trypanosoma uniporter Microbiology QR1-502 Miguel A. Chiurillo Noelia Lander Mayara S. Bertolini Melissa Storey Anibal E. Vercesi Roberto Docampo Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
description	ABSTRACT Trypanosoma cruzi is the agent of Chagas disease, and the finding that this parasite possesses a mitochondrial calcium uniporter (TcMCU) with characteristics similar to that of mammalian mitochondria was fundamental for the discovery of the molecular nature of MCU in eukaryotes. We report here that ablation of TcMCU, or its paralog TcMCUb, by clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 led to a marked decrease in mitochondrial Ca2+ uptake without affecting the membrane potential of these cells, whereas overexpression of each gene caused a significant increase in the ability of mitochondria to accumulate Ca2+. While TcMCU-knockout (KO) epimastigotes were viable and able to differentiate into trypomastigotes, infect host cells, and replicate normally, ablation of TcMCUb resulted in epimastigotes having an important growth defect, lower rates of respiration and metacyclogenesis, more pronounced autophagy changes under starvation, and significantly reduced infectivity. Overexpression of TcMCUb, in contrast to what was proposed for its mammalian ortholog, did not result in a dominant negative effect on TcMCU. IMPORTANCE The finding of a mitochondrial calcium uniporter (MCU) in Trypanosoma cruzi was essential for the discovery of the molecular nature of this transporter in mammals. In this work, we used the CRISPR/Cas9 technique that we recently developed for T. cruzi to knock out two components of the uniporter: MCU, the pore subunit, and MCUb, which was proposed as a negative regulator of MCU in human cells. In contrast to what occurs in human cells, MCU is not essential, while MCUb is essential for growth, differentiation, and infectivity; has a bioenergetic role; and does not act as a dominant negative subunit of MCU.
format	article
author	Miguel A. Chiurillo Noelia Lander Mayara S. Bertolini Melissa Storey Anibal E. Vercesi Roberto Docampo
author_facet	Miguel A. Chiurillo Noelia Lander Mayara S. Bertolini Melissa Storey Anibal E. Vercesi Roberto Docampo
author_sort	Miguel A. Chiurillo
title	Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
title_short	Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
title_full	Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
title_fullStr	Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
title_full_unstemmed	Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>
title_sort	different roles of mitochondrial calcium uniporter complex subunits in growth and infectivity of <italic toggle="yes">trypanosoma cruzi</italic>
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/2c66e00f1b9244cf888d9eea4da90625
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Different Roles of Mitochondrial Calcium Uniporter Complex Subunits in Growth and Infectivity of <italic toggle="yes">Trypanosoma cruzi</italic>

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