Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii
Toxoplasma gondii is a common zoonotic protozoan pathogen adapted to intracellular parasitism in many host cells of diverse organisms. Our previous work has identified 18 cyclic nucleotide phosphodiesterase (PDE) proteins encoded by the parasite genome, of which 11 are expressed during the lytic cyc...
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Elsevier
2022-01-01
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| Series: | Computational and Structural Biotechnology Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037022004275 |
| _version_ | 1797978171230912512 |
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| author | Kim Chi Vo Liberta Ruga Olympia Ekaterini Psathaki Rico Franzkoch Ute Distler Stefan Tenzer Michael Hensel Peter Hegemann Nishith Gupta |
| author_facet | Kim Chi Vo Liberta Ruga Olympia Ekaterini Psathaki Rico Franzkoch Ute Distler Stefan Tenzer Michael Hensel Peter Hegemann Nishith Gupta |
| author_sort | Kim Chi Vo |
| collection | DOAJ |
| description | Toxoplasma gondii is a common zoonotic protozoan pathogen adapted to intracellular parasitism in many host cells of diverse organisms. Our previous work has identified 18 cyclic nucleotide phosphodiesterase (PDE) proteins encoded by the parasite genome, of which 11 are expressed during the lytic cycle of its acutely-infectious tachyzoite stage in human cells. Here, we show that ten of these enzymes are promiscuous dual-specific phosphodiesterases, hydrolyzing cAMP and cGMP. TgPDE1 and TgPDE9, with a Km of 18 μM and 31 μM, respectively, are primed to hydrolyze cGMP, whereas TgPDE2 is highly specific to cAMP (Km, 14 μM). Immuno-electron microscopy revealed various subcellular distributions of TgPDE1, 2, and 9, including in the inner membrane complex, apical pole, plasma membrane, cytosol, dense granule, and rhoptry, indicating spatial control of signaling within tachyzoites. Notably, despite shared apical location and dual-catalysis, TgPDE8 and TgPDE9 are fully dispensable for the lytic cycle and show no functional redundancy. In contrast, TgPDE1 and TgPDE2 are individually required for optimal growth, and their collective loss is lethal to the parasite. In vitro phenotyping of these mutants revealed the roles of TgPDE1 and TgPDE2 in proliferation, gliding motility, invasion and egress of tachyzoites. Moreover, our enzyme inhibition assays in conjunction with chemogenetic phenotyping underpin TgPDE1 as a target of commonly-used PDE inhibitors, BIPPO and zaprinast. Finally, we identified a retinue of TgPDE1 and TgPDE2-interacting kinases and phosphatases, possibly regulating the enzymatic activity. In conclusion, our datasets on the catalytic function, physiological relevance, subcellular localization and drug inhibition of key phosphodiesterases highlight the previously-unanticipated plasticity and therapeutic potential of cyclic nucleotide signaling in T. gondii. |
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| format | Article |
| id | doaj.art-b52377f7bb7749cbacfacb575705260f |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-04-11T05:18:44Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
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| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-b52377f7bb7749cbacfacb575705260f2022-12-24T04:54:31ZengElsevierComputational and Structural Biotechnology Journal2001-03702022-01-012057755789Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondiiKim Chi Vo0Liberta Ruga1Olympia Ekaterini Psathaki2Rico Franzkoch3Ute Distler4Stefan Tenzer5Michael Hensel6Peter Hegemann7Nishith Gupta8Department of Molecular Parasitology, Institute of Biology, Faculty of Life Sciences, Humboldt University, Berlin, GermanyDepartment of Molecular Parasitology, Institute of Biology, Faculty of Life Sciences, Humboldt University, Berlin, GermanyUniversity of Osnabrück, Center of Cellular Nanoanalytics (CellNanOs), Integrated Bioimaging Faciltiy (iBiOs), GermanyUniversity of Osnabrück, Center of Cellular Nanoanalytics (CellNanOs), Integrated Bioimaging Faciltiy (iBiOs), GermanyInstitute of Immunology, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, GermanyInstitute of Immunology, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, GermanyUniversity of Osnabrück, Center of Cellular Nanoanalytics (CellNanOs), Integrated Bioimaging Faciltiy (iBiOs), GermanyDepartment of Molecular Parasitology, Institute of Biology, Faculty of Life Sciences, Humboldt University, Berlin, GermanyDepartment of Molecular Parasitology, Institute of Biology, Faculty of Life Sciences, Humboldt University, Berlin, Germany; Department of Biological Sciences, Birla Institute of Technology and Science, Pilani (BITS-P), Hyderabad, India; Corresponding author at: Institute of Biology, Faculty of Life Sciences, Humboldt University, Berlin, Germany.Toxoplasma gondii is a common zoonotic protozoan pathogen adapted to intracellular parasitism in many host cells of diverse organisms. Our previous work has identified 18 cyclic nucleotide phosphodiesterase (PDE) proteins encoded by the parasite genome, of which 11 are expressed during the lytic cycle of its acutely-infectious tachyzoite stage in human cells. Here, we show that ten of these enzymes are promiscuous dual-specific phosphodiesterases, hydrolyzing cAMP and cGMP. TgPDE1 and TgPDE9, with a Km of 18 μM and 31 μM, respectively, are primed to hydrolyze cGMP, whereas TgPDE2 is highly specific to cAMP (Km, 14 μM). Immuno-electron microscopy revealed various subcellular distributions of TgPDE1, 2, and 9, including in the inner membrane complex, apical pole, plasma membrane, cytosol, dense granule, and rhoptry, indicating spatial control of signaling within tachyzoites. Notably, despite shared apical location and dual-catalysis, TgPDE8 and TgPDE9 are fully dispensable for the lytic cycle and show no functional redundancy. In contrast, TgPDE1 and TgPDE2 are individually required for optimal growth, and their collective loss is lethal to the parasite. In vitro phenotyping of these mutants revealed the roles of TgPDE1 and TgPDE2 in proliferation, gliding motility, invasion and egress of tachyzoites. Moreover, our enzyme inhibition assays in conjunction with chemogenetic phenotyping underpin TgPDE1 as a target of commonly-used PDE inhibitors, BIPPO and zaprinast. Finally, we identified a retinue of TgPDE1 and TgPDE2-interacting kinases and phosphatases, possibly regulating the enzymatic activity. In conclusion, our datasets on the catalytic function, physiological relevance, subcellular localization and drug inhibition of key phosphodiesterases highlight the previously-unanticipated plasticity and therapeutic potential of cyclic nucleotide signaling in T. gondii.http://www.sciencedirect.com/science/article/pii/S2001037022004275ApicomplexaPhosphodiesterasecAMP & cGMP signalingLytic cycleTachyzoite |
| spellingShingle | Kim Chi Vo Liberta Ruga Olympia Ekaterini Psathaki Rico Franzkoch Ute Distler Stefan Tenzer Michael Hensel Peter Hegemann Nishith Gupta Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii Computational and Structural Biotechnology Journal Apicomplexa Phosphodiesterase cAMP & cGMP signaling Lytic cycle Tachyzoite |
| title | Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii |
| title_full | Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii |
| title_fullStr | Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii |
| title_full_unstemmed | Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii |
| title_short | Plasticity and therapeutic potential of cAMP and cGMP-specific phosphodiesterases in Toxoplasma gondii |
| title_sort | plasticity and therapeutic potential of camp and cgmp specific phosphodiesterases in toxoplasma gondii |
| topic | Apicomplexa Phosphodiesterase cAMP & cGMP signaling Lytic cycle Tachyzoite |
| url | http://www.sciencedirect.com/science/article/pii/S2001037022004275 |
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