A forward genetic screen identifies a negative regulator of rapid Ca2+-dependent cell egress (MS1) in the intracellular parasite Toxoplasma gondii
Toxoplasma gondii, like all apicomplexan parasites, uses Ca2+ signaling pathways to activate gliding motility to power tissue dissemination and host cell invasion and egress. A group of “plant-like” Ca2+-dependent protein kinases (CDPKs) transduces cytosolic Ca2+ flux into enzymatic activity, but ho...
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| Published in | The Journal of biological chemistry Vol. 292; no. 18; pp. 7662 - 7674 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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05.05.2017
American Society for Biochemistry and Molecular Biology |
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| Abstract | Toxoplasma gondii, like all apicomplexan parasites, uses Ca2+ signaling pathways to activate gliding motility to power tissue dissemination and host cell invasion and egress. A group of “plant-like” Ca2+-dependent protein kinases (CDPKs) transduces cytosolic Ca2+ flux into enzymatic activity, but how they function is poorly understood. To investigate how Ca2+ signaling activates egress through CDPKs, we performed a forward genetic screen to isolate gain-of-function mutants from an egress-deficient cdpk3 knockout strain. We recovered mutants that regained the ability to egress from host cells that harbored mutations in the gene Suppressor of Ca2+-dependent Egress 1 (SCE1). Global phosphoproteomic analysis showed that SCE1 deletion restored many Δcdpk3-dependent phosphorylation events to near wild-type levels. We also show that CDPK3-dependent SCE1 phosphorylation is required to relieve its suppressive activity to potentiate egress. In summary, our work has uncovered a novel component and suppressor of Ca2+-dependent cell egress during Toxoplasma lytic growth. |
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| AbstractList | Toxoplasma gondii, like all apicomplexan parasites, uses Ca2+ signaling pathways to activate gliding motility to power tissue dissemination and host cell invasion and egress. A group of “plant-like” Ca2+-dependent protein kinases (CDPKs) transduces cytosolic Ca2+ flux into enzymatic activity, but how they function is poorly understood. To investigate how Ca2+ signaling activates egress through CDPKs, we performed a forward genetic screen to isolate gain-of-function mutants from an egress-deficient cdpk3 knockout strain. We recovered mutants that regained the ability to egress from host cells that harbored mutations in the gene Suppressor of Ca2+-dependent Egress 1 (SCE1). Global phosphoproteomic analysis showed that SCE1 deletion restored many Δcdpk3-dependent phosphorylation events to near wild-type levels. We also show that CDPK3-dependent SCE1 phosphorylation is required to relieve its suppressive activity to potentiate egress. In summary, our work has uncovered a novel component and suppressor of Ca2+-dependent cell egress during Toxoplasma lytic growth. Toxoplasma gondii , like all apicomplexan parasites, uses Ca 2+ signaling pathways to activate gliding motility to power tissue dissemination and host cell invasion and egress. A group of “plant-like” Ca 2+ -dependent protein kinases (CDPKs) transduces cytosolic Ca 2+ flux into enzymatic activity, but how they function is poorly understood. To investigate how Ca 2+ signaling activates egress through CDPKs, we performed a forward genetic screen to isolate gain-of-function mutants from an egress-deficient cdpk3 knockout strain. We recovered mutants that regained the ability to egress from host cells that harbored mutations in the gene Suppressor of Ca 2+ -dependent Egress 1 (SCE1). Global phosphoproteomic analysis showed that SCE1 deletion restored many Δ cdpk3 -dependent phosphorylation events to near wild-type levels. We also show that CDPK3-dependent SCE1 phosphorylation is required to relieve its suppressive activity to potentiate egress. In summary, our work has uncovered a novel component and suppressor of Ca 2+ -dependent cell egress during Toxoplasma lytic growth. |
| Author | Stewart, Rebecca J. Tonkin, Christopher J. Schröder, Jan Lehane, Adele M. Papenfuss, Anthony T. Li, Dongdi Foster, Leonard J. McCoy, James M. Uboldi, Alessandro D. Scott, Nicollas E. |
| Author_xml | – sequence: 1 givenname: James M. surname: McCoy fullname: McCoy, James M. organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia – sequence: 2 givenname: Rebecca J. surname: Stewart fullname: Stewart, Rebecca J. organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia – sequence: 3 givenname: Alessandro D. surname: Uboldi fullname: Uboldi, Alessandro D. organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia – sequence: 4 givenname: Dongdi surname: Li fullname: Li, Dongdi organization: the Research School of Biology, Australian National University, Canberra, Australian Capital Territory 2601, Australia – sequence: 5 givenname: Jan surname: Schröder fullname: Schröder, Jan organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia – sequence: 6 givenname: Nicollas E. surname: Scott fullname: Scott, Nicollas E. organization: the University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada – sequence: 7 givenname: Anthony T. surname: Papenfuss fullname: Papenfuss, Anthony T. organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia – sequence: 8 givenname: Adele M. surname: Lehane fullname: Lehane, Adele M. organization: the Research School of Biology, Australian National University, Canberra, Australian Capital Territory 2601, Australia – sequence: 9 givenname: Leonard J. surname: Foster fullname: Foster, Leonard J. organization: the University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada – sequence: 10 givenname: Christopher J. surname: Tonkin fullname: Tonkin, Christopher J. email: tonkin@wehi.edu.au organization: From the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3052, Australia |
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| Issue | 18 |
| Keywords | molecular genetics parasite signaling Toxoplasma gondii proteomics apicomplexa forward genetic screen Ca2+ signalling host cell egress |
| Language | English |
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| SubjectTerms | apicomplexa Ca2+ signalling forward genetic screen host cell egress Microbiology molecular genetics parasite proteomics signaling Toxoplasma gondii |
| Title | A forward genetic screen identifies a negative regulator of rapid Ca2+-dependent cell egress (MS1) in the intracellular parasite Toxoplasma gondii |
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