Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>

ABSTRACT Toxoplasma gondii is a widespread intracellular pathogen infecting humans and a variety of animals. Previous studies have shown that Toxoplasma uses glucose and glutamine as the main carbon sources to support asexual reproduction, but neither nutrient is essential. Such metabolic flexibilit...

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Autores principales:	Ningbo Xia, Shu Ye, Xiaohan Liang, Pu Chen, Yanqin Zhou, Rui Fang, Junlong Zhao, Nishith Gupta, Shuzhen Yang, Jing Yuan, Bang Shen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Toxoplasma apicoplast lactate metabolic flexibility pyruvate kinase Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/64f32c2546cf4338a4b51309136ea13c
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spelling	oai:doaj.org-article:64f32c2546cf4338a4b51309136ea13c2021-11-15T15:55:24ZPyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>10.1128/mBio.00898-192150-7511https://doaj.org/article/64f32c2546cf4338a4b51309136ea13c2019-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00898-19https://doaj.org/toc/2150-7511ABSTRACT Toxoplasma gondii is a widespread intracellular pathogen infecting humans and a variety of animals. Previous studies have shown that Toxoplasma uses glucose and glutamine as the main carbon sources to support asexual reproduction, but neither nutrient is essential. Such metabolic flexibility may allow it to survive within diverse host cell types. Here, by focusing on the glycolytic enzyme pyruvate kinase (PYK) that converts phosphoenolpyruvate (PEP) into pyruvate, we found that Toxoplasma can also utilize lactate and alanine. We show that catabolism of all indicated carbon sources converges at pyruvate, and maintaining a constant pyruvate supply is critical to parasite growth. Toxoplasma expresses two PYKs: PYK1 in the cytosol and PYK2 in the apicoplast (a chloroplast relict). Genetic deletion of PYK2 did not noticeably affect parasite growth and virulence, which contrasts with the current model of carbon metabolism in the apicoplast. On the other hand, PYK1 was refractory to disruption. Conditional depletion of PYK1 resulted in global alteration of carbon metabolism, amylopectin accumulation, and reduced cellular ATP, leading to severe growth impairment. Notably, the attenuated growth of the PYK1-depleted mutant was partially rescued by lactate or alanine supplementation, and rescue by lactate required lactate dehydrogenase activity to convert it to pyruvate. Moreover, depletion of PYK1 in conjunction with PYK2 ablation led to accentuated loss of apicoplasts and complete growth arrest. Together, our results underline a critical role of pyruvate homeostasis in determining the metabolic flexibility and apicoplast maintenance, and they significantly extend our current understanding of carbon metabolism in T. gondii. IMPORTANCE Toxoplasma gondii infects almost all warm-blooded animals, and metabolic flexibility is deemed critical for its successful parasitism in diverse hosts. Glucose and glutamine are the major carbon sources to support parasite growth. In this study, we found that Toxoplasma is also competent in utilizing lactate and alanine and, thus, exhibits exceptional metabolic versatility. Notably, all these nutrients need to be converted to pyruvate to fuel the lytic cycle, and achieving a continued pyruvate supply is vital to parasite survival and metabolic flexibility. Although pyruvate can be generated by two distinct pyruvate kinases, located in cytosol and apicoplast, respectively, the cytosolic enzyme is the main source of subcellular pyruvate, and cooperative usage of pyruvate among multiple organelles is critical for parasite growth and virulence. These findings expand our current understanding of carbon metabolism in Toxoplasma gondii and related parasites while providing a basis for designing novel antiparasitic interventions.Ningbo XiaShu YeXiaohan LiangPu ChenYanqin ZhouRui FangJunlong ZhaoNishith GuptaShuzhen YangJing YuanBang ShenAmerican Society for MicrobiologyarticleToxoplasmaapicoplastlactatemetabolic flexibilitypyruvate kinaseMicrobiologyQR1-502ENmBio, Vol 10, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Toxoplasma apicoplast lactate metabolic flexibility pyruvate kinase Microbiology QR1-502
spellingShingle	Toxoplasma apicoplast lactate metabolic flexibility pyruvate kinase Microbiology QR1-502 Ningbo Xia Shu Ye Xiaohan Liang Pu Chen Yanqin Zhou Rui Fang Junlong Zhao Nishith Gupta Shuzhen Yang Jing Yuan Bang Shen Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
description	ABSTRACT Toxoplasma gondii is a widespread intracellular pathogen infecting humans and a variety of animals. Previous studies have shown that Toxoplasma uses glucose and glutamine as the main carbon sources to support asexual reproduction, but neither nutrient is essential. Such metabolic flexibility may allow it to survive within diverse host cell types. Here, by focusing on the glycolytic enzyme pyruvate kinase (PYK) that converts phosphoenolpyruvate (PEP) into pyruvate, we found that Toxoplasma can also utilize lactate and alanine. We show that catabolism of all indicated carbon sources converges at pyruvate, and maintaining a constant pyruvate supply is critical to parasite growth. Toxoplasma expresses two PYKs: PYK1 in the cytosol and PYK2 in the apicoplast (a chloroplast relict). Genetic deletion of PYK2 did not noticeably affect parasite growth and virulence, which contrasts with the current model of carbon metabolism in the apicoplast. On the other hand, PYK1 was refractory to disruption. Conditional depletion of PYK1 resulted in global alteration of carbon metabolism, amylopectin accumulation, and reduced cellular ATP, leading to severe growth impairment. Notably, the attenuated growth of the PYK1-depleted mutant was partially rescued by lactate or alanine supplementation, and rescue by lactate required lactate dehydrogenase activity to convert it to pyruvate. Moreover, depletion of PYK1 in conjunction with PYK2 ablation led to accentuated loss of apicoplasts and complete growth arrest. Together, our results underline a critical role of pyruvate homeostasis in determining the metabolic flexibility and apicoplast maintenance, and they significantly extend our current understanding of carbon metabolism in T. gondii. IMPORTANCE Toxoplasma gondii infects almost all warm-blooded animals, and metabolic flexibility is deemed critical for its successful parasitism in diverse hosts. Glucose and glutamine are the major carbon sources to support parasite growth. In this study, we found that Toxoplasma is also competent in utilizing lactate and alanine and, thus, exhibits exceptional metabolic versatility. Notably, all these nutrients need to be converted to pyruvate to fuel the lytic cycle, and achieving a continued pyruvate supply is vital to parasite survival and metabolic flexibility. Although pyruvate can be generated by two distinct pyruvate kinases, located in cytosol and apicoplast, respectively, the cytosolic enzyme is the main source of subcellular pyruvate, and cooperative usage of pyruvate among multiple organelles is critical for parasite growth and virulence. These findings expand our current understanding of carbon metabolism in Toxoplasma gondii and related parasites while providing a basis for designing novel antiparasitic interventions.
format	article
author	Ningbo Xia Shu Ye Xiaohan Liang Pu Chen Yanqin Zhou Rui Fang Junlong Zhao Nishith Gupta Shuzhen Yang Jing Yuan Bang Shen
author_facet	Ningbo Xia Shu Ye Xiaohan Liang Pu Chen Yanqin Zhou Rui Fang Junlong Zhao Nishith Gupta Shuzhen Yang Jing Yuan Bang Shen
author_sort	Ningbo Xia
title	Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_short	Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full	Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_fullStr	Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full_unstemmed	Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_sort	pyruvate homeostasis as a determinant of parasite growth and metabolic plasticity in <named-content content-type="genus-species">toxoplasma gondii</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/64f32c2546cf4338a4b51309136ea13c
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Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in <named-content content-type="genus-species">Toxoplasma gondii</named-content>

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