A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages

Yifan Wang; Lamba Omar Sangare; Tatiana C.  Paredes-Santos; Shruthi  Krishnamurthy; Musa Hassan; Anna M.  Furuta; Benedikt M.  Markus; Sebastian  Lourido; Jeroen P.J.  Saeij

doi:10.1101/867705

A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages

Yifan Wang, Lamba Omar Sangare, Tatiana C. Paredes-Santos, Shruthi Krishnamurthy, Musa Hassan, Anna M. Furuta, Benedikt M. Markus, Sebastian Lourido, Jeroen P.J. Saeij

Research output: Working paper

Abstract / Description of output

Macrophages play an essential role in the early immune response against Toxoplasma and are the cell type preferentially infected by the parasite in vivo. Interferon gamma (IFNγ) elicits a variety of anti-Toxoplasma activities in macrophages. Using a genome-wide CRISPR screen we identified ∼130 Toxoplasma genes that determine parasite fitness in naїve macrophages and ∼466 genes that determine fitness in IFNγ-stimulated murine macrophages, seven of which we investigated and confirmed. We show that one of these genes encodes dense granule protein GRA45, which contains a putative chaperone-like domain, and which we show is critical in preventing other GRA effectors from aggregating. Parasites lacking GRA45 mislocalize GRA effectors upon secretion, are more susceptible to IFNγ-mediated growth inhibition, and have reduced virulence in mice. Our results provide a resource for the community to further explore the function of Toxoplasma genes that determine fitness in IFNγ-stimulated macrophages.

Original language	English
Publisher	bioRxiv, at Cold Spring Harbor Laboratory
DOIs	https://doi.org/10.1101/867705
Publication status	Published - 6 Dec 2019

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Name	bioRxiv
Publisher	bioRxiv, at Cold Spring Harbor Laboratory

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10.1101/867705

A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages
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Wang, Y., Sangare, L. O., Paredes-Santos, T. C., Krishnamurthy, S., Hassan, M., Furuta, A. M., Markus, B. M., Lourido, S., & Saeij, J. P. J. (2019). A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages. (bioRxiv). bioRxiv, at Cold Spring Harbor Laboratory. https://doi.org/10.1101/867705

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title = "A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages",

abstract = "Macrophages play an essential role in the early immune response against Toxoplasma and are the cell type preferentially infected by the parasite in vivo. Interferon gamma (IFNγ) elicits a variety of anti-Toxoplasma activities in macrophages. Using a genome-wide CRISPR screen we identified ∼130 Toxoplasma genes that determine parasite fitness in naїve macrophages and ∼466 genes that determine fitness in IFNγ-stimulated murine macrophages, seven of which we investigated and confirmed. We show that one of these genes encodes dense granule protein GRA45, which contains a putative chaperone-like domain, and which we show is critical in preventing other GRA effectors from aggregating. Parasites lacking GRA45 mislocalize GRA effectors upon secretion, are more susceptible to IFNγ-mediated growth inhibition, and have reduced virulence in mice. Our results provide a resource for the community to further explore the function of Toxoplasma genes that determine fitness in IFNγ-stimulated macrophages.",

author = "Yifan Wang and Sangare, {Lamba Omar} and Paredes-Santos, {Tatiana C.} and Shruthi Krishnamurthy and Musa Hassan and Furuta, {Anna M.} and Markus, {Benedikt M.} and Sebastian Lourido and Saeij, {Jeroen P.J.}",

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doi = "10.1101/867705",

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AU - Sangare, Lamba Omar

AU - Paredes-Santos, Tatiana C.

AU - Krishnamurthy, Shruthi

AU - Hassan, Musa

AU - Furuta, Anna M.

AU - Markus, Benedikt M.

AU - Lourido, Sebastian

AU - Saeij, Jeroen P.J.

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N2 - Macrophages play an essential role in the early immune response against Toxoplasma and are the cell type preferentially infected by the parasite in vivo. Interferon gamma (IFNγ) elicits a variety of anti-Toxoplasma activities in macrophages. Using a genome-wide CRISPR screen we identified ∼130 Toxoplasma genes that determine parasite fitness in naїve macrophages and ∼466 genes that determine fitness in IFNγ-stimulated murine macrophages, seven of which we investigated and confirmed. We show that one of these genes encodes dense granule protein GRA45, which contains a putative chaperone-like domain, and which we show is critical in preventing other GRA effectors from aggregating. Parasites lacking GRA45 mislocalize GRA effectors upon secretion, are more susceptible to IFNγ-mediated growth inhibition, and have reduced virulence in mice. Our results provide a resource for the community to further explore the function of Toxoplasma genes that determine fitness in IFNγ-stimulated macrophages.

AB - Macrophages play an essential role in the early immune response against Toxoplasma and are the cell type preferentially infected by the parasite in vivo. Interferon gamma (IFNγ) elicits a variety of anti-Toxoplasma activities in macrophages. Using a genome-wide CRISPR screen we identified ∼130 Toxoplasma genes that determine parasite fitness in naїve macrophages and ∼466 genes that determine fitness in IFNγ-stimulated murine macrophages, seven of which we investigated and confirmed. We show that one of these genes encodes dense granule protein GRA45, which contains a putative chaperone-like domain, and which we show is critical in preventing other GRA effectors from aggregating. Parasites lacking GRA45 mislocalize GRA effectors upon secretion, are more susceptible to IFNγ-mediated growth inhibition, and have reduced virulence in mice. Our results provide a resource for the community to further explore the function of Toxoplasma genes that determine fitness in IFNγ-stimulated macrophages.

U2 - 10.1101/867705

DO - 10.1101/867705

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BT - A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages

PB - bioRxiv, at Cold Spring Harbor Laboratory

ER -

A genome-wide loss-of-function screen identifies Toxoplasma gondii genes that determine fitness in interferon gamma-activated murine macrophages

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