Proximity biotinylation reveals novel secreted dense granule proteins of Toxoplasma gondii bradyzoites.

Autor: Nadipuram SM; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, United States of America.; Division of Pediatric Infectious Diseases, Department of Pediatrics, Cedar-Sinai Medical Center, Los Angeles, California, United States of America., Thind AC; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, United States of America.; Molecular Biology Institute, University of California, Los Angeles, California, United States of America., Rayatpisheh S; Department of Biological Chemistry and Institute of Genomics and Proteomics, University of California, Los Angeles, California, United States of America., Wohlschlegel JA; Molecular Biology Institute, University of California, Los Angeles, California, United States of America.; Department of Biological Chemistry and Institute of Genomics and Proteomics, University of California, Los Angeles, California, United States of America., Bradley PJ; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, United States of America.; Molecular Biology Institute, University of California, Los Angeles, California, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2020 May 06; Vol. 15 (5), pp. e0232552. Date of Electronic Publication: 2020 May 06 (Print Publication: 2020).
DOI: 10.1371/journal.pone.0232552
Abstrakt: Toxoplasma gondii is an obligate intracellular parasite which is capable of establishing life-long chronic infection in any mammalian host. During the intracellular life cycle, the parasite secretes an array of proteins into the parasitophorous vacuole (PV) where it resides. Specialized organelles called the dense granules secrete GRA proteins that are known to participate in nutrient acquisition, immune evasion, and host cell-cycle manipulation. Although many GRAs have been discovered which are expressed during the acute infection mediated by tachyzoites, little is known about those that participate in the chronic infection mediated by the bradyzoite form of the parasite. In this study, we sought to uncover novel bradyzoite-upregulated GRA proteins using proximity biotinylation, which we previously used to examine the secreted proteome of the tachyzoites. Using a fusion of the bradyzoite upregulated protein MAG1 to BirA* as bait and a strain with improved switch efficiency, we identified a number of novel GRA proteins which are expressed in bradyzoites. After using the CRISPR/Cas9 system to characterize these proteins by gene knockout, we focused on one of these GRAs (GRA55) and found it was important for the establishment or maintenance of cysts in the mouse brain. These findings highlight new components of the GRA proteome of the tissue-cyst life stage of T. gondii and identify potential targets that are important for maintenance of parasite persistence in vivo.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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