TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>

ABSTRACT Toxoplasma gondii is a widespread protozoan parasite that causes potentially life-threatening opportunistic disease. New inhibitors of parasite replication are urgently needed, as the current antifolate treatment is also toxic to patients. Microtubules are essential cytoskeletal components...

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Autores principales:	Joseph M. Varberg, Leah R. Padgett, Gustavo Arrizabalaga, William J. Sullivan
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	microtubules cytoskeleton lysine acetylation Mec-17 acetyltransferase endodyogeny Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b6ab5c8ac25544958db494489944b7ca
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spelling	oai:doaj.org-article:b6ab5c8ac25544958db494489944b7ca2021-11-15T15:21:38ZTgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>10.1128/mSphere.00088-152379-5042https://doaj.org/article/b6ab5c8ac25544958db494489944b7ca2016-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00088-15https://doaj.org/toc/2379-5042ABSTRACT Toxoplasma gondii is a widespread protozoan parasite that causes potentially life-threatening opportunistic disease. New inhibitors of parasite replication are urgently needed, as the current antifolate treatment is also toxic to patients. Microtubules are essential cytoskeletal components that have been selectively targeted in microbial pathogens; further study of tubulin in Toxoplasma may reveal novel therapeutic opportunities. It has been noted that α-tubulin acetylation at lysine 40 (K40) is enriched during daughter parasite formation, but the impact of this modification on Toxoplasma division and the enzyme mediating its delivery have not been identified. We performed mutational analyses to provide evidence that K40 acetylation stabilizes Toxoplasma microtubules and is required for parasite replication. We also show that an unusual Toxoplasma homologue of α-tubulin acetyltransferase (TgATAT) is expressed in a cell cycle-regulated manner and that its expression peaks during division. Disruption of TgATAT with CRISPR/Cas9 ablates K40 acetylation and induces replication defects; parasites appear to initiate mitosis yet exhibit incomplete or improper nuclear division. Together, these findings establish the importance of tubulin acetylation, exposing a new vulnerability in Toxoplasma that could be pharmacologically targeted. IMPORTANCE Toxoplasma gondii is an opportunistic parasite that infects at least one-third of the world population. New treatments for the disease (toxoplasmosis) are needed since current drugs are toxic to patients. Microtubules are essential cellular structures built from tubulin that show promise as antimicrobial drug targets. Microtubules can be regulated by chemical modification, such as acetylation on lysine 40 (K40). To determine the role of K40 acetylation in Toxoplasma and whether it is a liability to the parasite, we performed mutational analyses of the α-tubulin gene. Our results indicate that parasites cannot survive without K40 acetylation unless microtubules are stabilized with a secondary mutation. Additionally, we identified the parasite enzyme that acetylates α-tubulin (TgATAT). Genetic disruption of TgATAT caused severe defects in parasite replication, further highlighting the importance of α-tubulin K40 acetylation in Toxoplasma and its promise as a potential new drug target.Joseph M. VarbergLeah R. PadgettGustavo ArrizabalagaWilliam J. SullivanAmerican Society for Microbiologyarticlemicrotubulescytoskeletonlysine acetylationMec-17acetyltransferaseendodyogenyMicrobiologyQR1-502ENmSphere, Vol 1, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	microtubules cytoskeleton lysine acetylation Mec-17 acetyltransferase endodyogeny Microbiology QR1-502
spellingShingle	microtubules cytoskeleton lysine acetylation Mec-17 acetyltransferase endodyogeny Microbiology QR1-502 Joseph M. Varberg Leah R. Padgett Gustavo Arrizabalaga William J. Sullivan TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
description	ABSTRACT Toxoplasma gondii is a widespread protozoan parasite that causes potentially life-threatening opportunistic disease. New inhibitors of parasite replication are urgently needed, as the current antifolate treatment is also toxic to patients. Microtubules are essential cytoskeletal components that have been selectively targeted in microbial pathogens; further study of tubulin in Toxoplasma may reveal novel therapeutic opportunities. It has been noted that α-tubulin acetylation at lysine 40 (K40) is enriched during daughter parasite formation, but the impact of this modification on Toxoplasma division and the enzyme mediating its delivery have not been identified. We performed mutational analyses to provide evidence that K40 acetylation stabilizes Toxoplasma microtubules and is required for parasite replication. We also show that an unusual Toxoplasma homologue of α-tubulin acetyltransferase (TgATAT) is expressed in a cell cycle-regulated manner and that its expression peaks during division. Disruption of TgATAT with CRISPR/Cas9 ablates K40 acetylation and induces replication defects; parasites appear to initiate mitosis yet exhibit incomplete or improper nuclear division. Together, these findings establish the importance of tubulin acetylation, exposing a new vulnerability in Toxoplasma that could be pharmacologically targeted. IMPORTANCE Toxoplasma gondii is an opportunistic parasite that infects at least one-third of the world population. New treatments for the disease (toxoplasmosis) are needed since current drugs are toxic to patients. Microtubules are essential cellular structures built from tubulin that show promise as antimicrobial drug targets. Microtubules can be regulated by chemical modification, such as acetylation on lysine 40 (K40). To determine the role of K40 acetylation in Toxoplasma and whether it is a liability to the parasite, we performed mutational analyses of the α-tubulin gene. Our results indicate that parasites cannot survive without K40 acetylation unless microtubules are stabilized with a secondary mutation. Additionally, we identified the parasite enzyme that acetylates α-tubulin (TgATAT). Genetic disruption of TgATAT caused severe defects in parasite replication, further highlighting the importance of α-tubulin K40 acetylation in Toxoplasma and its promise as a potential new drug target.
format	article
author	Joseph M. Varberg Leah R. Padgett Gustavo Arrizabalaga William J. Sullivan
author_facet	Joseph M. Varberg Leah R. Padgett Gustavo Arrizabalaga William J. Sullivan
author_sort	Joseph M. Varberg
title	TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_short	TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full	TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_fullStr	TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full_unstemmed	TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_sort	tgatat-mediated α-tubulin acetylation is required for division of the protozoan parasite <named-content content-type="genus-species">toxoplasma gondii</named-content>
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/b6ab5c8ac25544958db494489944b7ca
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TgATAT-Mediated α-Tubulin Acetylation Is Required for Division of the Protozoan Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>

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