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Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma

Toxoplasma gondii is a parasite that replicates within a specialized compartment called the parasitophorous vacuole (PV), which is surrounded by the PV membrane (PVM). To obtain essential nutrients, Toxoplasma must transport molecules across the PVM, a process mediated by the secreted parasite prote...

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Autores principales: Paredes-Santos, Tatiana C., Bitew, Mebratu A., Swale, Christopher, Rodriguez, Felipe, Krishnamurthy, Shruthi, Wang, Yifan, Maru, Parag, Sangaré, Lamba Omar, Saeij, Jeroen P. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10409377/
https://www.ncbi.nlm.nih.gov/pubmed/37498952
http://dx.doi.org/10.1371/journal.ppat.1011543
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author Paredes-Santos, Tatiana C.
Bitew, Mebratu A.
Swale, Christopher
Rodriguez, Felipe
Krishnamurthy, Shruthi
Wang, Yifan
Maru, Parag
Sangaré, Lamba Omar
Saeij, Jeroen P. J.
author_facet Paredes-Santos, Tatiana C.
Bitew, Mebratu A.
Swale, Christopher
Rodriguez, Felipe
Krishnamurthy, Shruthi
Wang, Yifan
Maru, Parag
Sangaré, Lamba Omar
Saeij, Jeroen P. J.
author_sort Paredes-Santos, Tatiana C.
collection PubMed
description Toxoplasma gondii is a parasite that replicates within a specialized compartment called the parasitophorous vacuole (PV), which is surrounded by the PV membrane (PVM). To obtain essential nutrients, Toxoplasma must transport molecules across the PVM, a process mediated by the secreted parasite proteins GRA17 and GRA23. These proteins form pores in the PVM through which small molecules can diffuse in and out of the PV. GRA17 and GRA23 are synthetically lethal, suggesting that at least one pore type is essential for parasite survival. In the ‘nutrient sensitized’ Δgra17 strain it is likely that other Toxoplasma genes become essential, because they mediate nutrient acquisition from the host or are involved in the trafficking of GRA23 to the PVM. To identify these genes, a genome-wide loss-of-function screen was performed in wild-type and Δgra17 parasites, which identified multiple genes that were synthetically sick/lethal with GRA17. Several of these genes were involved in the correct localization of GRAs, including GRA17/GRA23, to the PVM. One of the top hits, GRA72, was predicted to form a pore on the PVM, and its deletion led to the formation of enlarged “bubble vacuoles” with reduced PVM small molecule permeability, similar to what was previously observed for Δgra17 parasites. Furthermore, Δgra72 parasites had reduced in vitro growth and virulence in mice. These findings suggest that in the absence of GRA17, other genes become essential, likely because they play a role in the proper localization of GRA23 (and other GRAs) or because they determine host-derived nutrient acquisition at the PVM.
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spelling pubmed-104093772023-08-09 Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma Paredes-Santos, Tatiana C. Bitew, Mebratu A. Swale, Christopher Rodriguez, Felipe Krishnamurthy, Shruthi Wang, Yifan Maru, Parag Sangaré, Lamba Omar Saeij, Jeroen P. J. PLoS Pathog Research Article Toxoplasma gondii is a parasite that replicates within a specialized compartment called the parasitophorous vacuole (PV), which is surrounded by the PV membrane (PVM). To obtain essential nutrients, Toxoplasma must transport molecules across the PVM, a process mediated by the secreted parasite proteins GRA17 and GRA23. These proteins form pores in the PVM through which small molecules can diffuse in and out of the PV. GRA17 and GRA23 are synthetically lethal, suggesting that at least one pore type is essential for parasite survival. In the ‘nutrient sensitized’ Δgra17 strain it is likely that other Toxoplasma genes become essential, because they mediate nutrient acquisition from the host or are involved in the trafficking of GRA23 to the PVM. To identify these genes, a genome-wide loss-of-function screen was performed in wild-type and Δgra17 parasites, which identified multiple genes that were synthetically sick/lethal with GRA17. Several of these genes were involved in the correct localization of GRAs, including GRA17/GRA23, to the PVM. One of the top hits, GRA72, was predicted to form a pore on the PVM, and its deletion led to the formation of enlarged “bubble vacuoles” with reduced PVM small molecule permeability, similar to what was previously observed for Δgra17 parasites. Furthermore, Δgra72 parasites had reduced in vitro growth and virulence in mice. These findings suggest that in the absence of GRA17, other genes become essential, likely because they play a role in the proper localization of GRA23 (and other GRAs) or because they determine host-derived nutrient acquisition at the PVM. Public Library of Science 2023-07-27 /pmc/articles/PMC10409377/ /pubmed/37498952 http://dx.doi.org/10.1371/journal.ppat.1011543 Text en © 2023 Paredes-Santos et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Paredes-Santos, Tatiana C.
Bitew, Mebratu A.
Swale, Christopher
Rodriguez, Felipe
Krishnamurthy, Shruthi
Wang, Yifan
Maru, Parag
Sangaré, Lamba Omar
Saeij, Jeroen P. J.
Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title_full Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title_fullStr Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title_full_unstemmed Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title_short Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma
title_sort genome-wide crispr screen identifies genes synthetically lethal with gra17, a nutrient channel encoding gene in toxoplasma
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10409377/
https://www.ncbi.nlm.nih.gov/pubmed/37498952
http://dx.doi.org/10.1371/journal.ppat.1011543
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