A Toxoplasma gondii patatin-like phospholipase contributes to host cell invasion
| Autor: | Lindsey L. Koch, Naomi S. Morrissette, Sarah K. Wilson, Justine Heckendorn, Maryse Lebrun, Laura J. Knoll, Bruno Martorelli Di Genova, Peggy J. Rooney |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Physiology
Hydrolases Mutant Complement System Protozoan Proteins Biochemistry Toxoplasma Gondii Mice Immune Physiology Medicine and Health Sciences Serine Amino Acids Biology (General) Protozoans 0303 health sciences Immune System Proteins Virulence Organic Compounds 030302 biochemistry & molecular biology Esterases Eukaryota Animal Models Enzymes Complementation Chemistry Patatin-like phospholipase Experimental Organism Systems Phospholipases Physical Sciences Toxoplasma Toxoplasmosis Research Article Virulence Factors QH301-705.5 Immunology Mouse Models Biology Research and Analysis Methods Microbiology 03 medical and health sciences Model Organisms Virology Hydroxyl Amino Acids Genetics Parasitic Diseases Animals Secretion Molecular Biology 030304 developmental biology Rhoptry Intracellular parasite Host Cells Organic Chemistry Wild type Organisms Chemical Compounds Toxoplasma gondii Biology and Life Sciences Proteins RC581-607 biology.organism_classification Parasitic Protozoans Immune System Animal Studies Enzymology Parasitology Immunologic diseases. Allergy Physiological Processes Viral Transmission and Infection |
| Zdroj: | PLoS Pathogens, Vol 16, Iss 7, p e1008650 (2020) PLoS Pathogens |
| ISSN: | 1553-7374 1553-7366 |
| Popis: | Toxoplasma gondii is an obligate intracellular parasite that can invade any nucleated cell of any warm-blooded animal. In a previous screen to identify virulence determinants, disruption of gene TgME49_305140 generated a T. gondii mutant that could not establish a chronic infection in mice. The protein product of TgME49_305140, here named TgPL3, is a 277 kDa protein with a patatin-like phospholipase (PLP) domain and a microtubule binding domain. Antibodies generated against TgPL3 show that it is localized to the apical cap. Using a rapid selection FACS-based CRISPR/Cas-9 method, a TgPL3 deletion strain (ΔTgPL3) was generated. ΔTgPL3 parasites have defects in host cell invasion, which may be caused by reduced rhoptry secretion. We generated complementation clones with either wild type TgPL3 or an active site mutation in the PLP domain by converting the catalytic serine to an alanine, ΔTgPL3::TgPL3S1409A (S1409A). Complementation of ΔTgPL3 with wild type TgPL3 restored all phenotypes, while S1409A did not, suggesting that phospholipase activity is necessary for these phenotypes. ΔTgPL3 and S1409A parasites are also virtually avirulent in vivo but induce a robust antibody response. Vaccination with ΔTgPL3 and S1409A parasites protected mice against subsequent challenge with a lethal dose of Type I T. gondii parasites, making ΔTgPL3 a compelling vaccine candidate. These results demonstrate that TgPL3 has a role in rhoptry secretion, host cell invasion and survival of T. gondii during acute mouse infection. Author summary Toxoplasma gondii is a eukaryotic parasite commonly found in warm-blooded animals worldwide. Successful replication of T. gondii within the host relies on its sophisticated cell invasion mechanism. Previous studies found that disruption of the T. gondii gene TgME49_305140 generated a mutant that could not establish a chronic infection in mice. The protein product of this gene, named TgPL3, is large with both patatin-like phospholipase and microtubule binding domains. Here we show that TgPL3 is localized to the apical end of T. gondii, which is used for invasion. A mutant T. gondii strain with a deletion of TgPL3 (ΔTgPL3) was generated. ΔTgPL3 parasites are defective for host cell invasion and do not cause disease in mice, even at high doses. Complementation of the TgPL3 gene back into ΔTgPL3 parasites repairs the invasion and mice infection defects. Moreover, complementation of ΔTgPL3 with a point mutation in the active site of the phospholipase domain (S1409A) did not rescue the invasion and mouse infection defects, suggesting the PLP domain is responsible for these phenotypes. Vaccination of mice with ΔTgPL3 and S1409A parasites protected them against lethal challenge with T. gondii, highlighting the potential of ΔTgPL3 as a vaccine strain. |
| Databáze: | OpenAIRE |
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