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Title: A Novel Secreted Protein, MYR1, Is Central to Toxoplasma ’s Manipulation of Host Cells

Journal Article · · mBio (Online)
 [1];  [2];  [2];  [3];  [2];  [2];  [4];  [4];  [3];  [2]
  1. Stanford Univ. School of Medicine, CA (United States); Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
  2. Stanford Univ. School of Medicine, CA (United States)
  3. Univ. of California, Santa Cruz, CA (United States)
  4. Washington Univ., St. Louis, MO (United States)

ABSTRACT The intracellular protozoanToxoplasma gondiidramatically reprograms the transcriptome of host cells it infects, including substantially up-regulating the host oncogene c-myc. By applying a flow cytometry-based selection to infected mouse cells expressing green fluorescent protein fused to c-Myc (c-Myc–GFP), we isolated mutant tachyzoites defective in this host c-Myc up-regulation. Whole-genome sequencing of three such mutants led to the identification ofMYR1(Mycregulation1;TGGT1_254470) as essential for c-Myc induction. MYR1 is a secreted protein that requires TgASP5 to be cleaved into two stable portions, both of which are ultimately found within the parasitophorous vacuole and at the parasitophorous vacuole membrane. Deletion ofMYR1revealed that in addition to its requirement for c-Myc up-regulation, the MYR1 protein is needed for the ability ofToxoplasmatachyzoites to modulate several other important host pathways, including those mediated by the dense granule effectors GRA16 and GRA24. This result, combined with its location at the parasitophorous vacuole membrane, suggested that MYR1 might be a component of the machinery that translocatesToxoplasmaeffectors from the parasitophorous vacuole into the host cytosol. Support for this possibility was obtained by showing that transit of GRA24 to the host nucleus is indeed MYR1-dependent. As predicted by this pleiotropic phenotype, parasites deficient inMYR1were found to be severely attenuated in a mouse model of infection. We conclude, therefore, that MYR1 is a novel protein that plays a critical role in howToxoplasmadelivers effector proteins to the infected host cell and that this is crucial to virulence. IMPORTANCEToxoplasma gondiiis an important human pathogen and a model for the study of intracellular parasitism. Infection of the host cell withToxoplasmatachyzoites involves the introduction of protein effectors, including many that are initially secreted into the parasitophorous vacuole but must ultimately translocate to the host cell cytosol to function. The work reported here identified a novel protein that is required for this translocation. These results give new insight into a very unusual cell biology process as well as providing a potential handle on a pathway that is necessary for virulence and, therefore, a new potential target for chemotherapy.

Research Organization:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Sponsoring Organization:
USDOE; National Institutes of Health (NIH)
Grant/Contract Number:
R21-AI112962; RO1-AI73756; T32-AI007328; P01-35HG000205; T32-AI007290; F31-AI120649; K08AI102946-01; S10RR025518-01; P30-NS069375; AC52-07NA27344
OSTI ID:
1259517
Alternate ID(s):
OSTI ID: 1420295
Report Number(s):
LLNL-JRNL-736669
Journal Information:
mBio (Online), Vol. 7, Issue 1; ISSN 2150-7511
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 97 works
Citation information provided by
Web of Science

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Cited By (32)

MYR1-Dependent Effectors Are the Major Drivers of a Host Cell’s Early Response to Toxoplasma , Including Counteracting MYR1-Independent Effects journal April 2018
Recent advances in understanding apicomplexan parasites journal January 2016
Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in Toxoplasma- Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase journal July 2019
Toxoplasma gondii Parasitophorous Vacuole Membrane-Associated Dense Granule Proteins Orchestrate Chronic Infection and GRA12 Underpins Resistance to Host Gamma Interferon journal July 2019
Toxoplasma gondii Parasitophorous Vacuole Membrane-Associated Dense Granule Proteins Regulate Maturation of the Cyst Wall journal January 2020
An in vitro model of intestinal infection reveals a developmentally regulated transcriptome of Toxoplasma sporozoites and a NF-κB-like signature in infected host cells journal March 2017
Enrichment and Proteomic Characterization of the Cyst Wall from In Vitro Toxoplasma gondii Cysts journal April 2019
Coimmunoprecipitation with MYR1 Identifies Three Additional Proteins within the Toxoplasma gondii Parasitophorous Vacuole Required for Translocation of Dense Granule Effectors into Host Cells journal February 2020
Human Placental Syncytiotrophoblasts Restrict Toxoplasma gondii Attachment and Replication and Respond to Infection by Producing Immunomodulatory Chemokines journal January 2018
A divergent kinase lacking the glycine-rich loop regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole posted_content August 2018
A CRISPR platform for targeted in vivo screens identifies Toxoplasma gondii virulence factors in mice. text January 2020
The Secreted Acid Phosphatase Domain-Containing GRA44 from Toxoplasma gondii Is Required for c-Myc Induction in Infected Cells journal February 2020
Toxoplasma Does Not Secrete the GRA16 and GRA24 Effectors Beyond the Parasitophorous Vacuole Membrane of Tissue Cysts journal October 2018
Divergent kinase regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole journal March 2019
Naïve CD8 T cell IFNγ responses to a vacuolar antigen are regulated by an inflammasome-independent NLRP3 pathway and Toxoplasma gondii ROP5 journal January 2020
Toxoplasma gondii ROP16 kinase silences the cyclin B1 gene promoter by hijacking host cell UHRF1-dependent epigenetic pathways journal September 2019
Aspartyl Protease 5 matures virulence factors found at the host-parasite interface in Toxoplasma gondii journal February 2018
Identification of a novel protein complex essential for effector translocation across the parasitophorous vacuole membrane of Toxoplasma gondii journal January 2018
Toxoplasma Effectors Targeting Host Signaling and Transcription journal April 2017
In vitro characterization of protein effector export in the bradyzoite stage of Toxoplasma gondii journal January 2020
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Characterization of a Toxoplasma effector uncovers an alternative GSK3/β-catenin-regulatory pathway of inflammation journal October 2018
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A CRISPR platform for targeted in vivo screens identifies Toxoplasma gondii virulence factors in mice. text January 2020
Divergent kinase regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole text January 2019
A CRISPR platform for targeted in vivo screens identifies Toxoplasma gondii virulence factors in mice journal September 2019
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Congenital Transmission of Toxoplasma gondii After Experimental Reinfection With Brazilian Typical Strains in Chronically Infected Sheep journal April 2019

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