Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1

Matthew A. Child, Megan Garland, Ian Foe, Peter Madzelan, Moritz Treeck, Wouter A. Van Der Linden, Kristina Oresic Bender, Eranthie Weerapana, Mark A. Wilson, John C. Boothroyd, Michael L. Reese, Matthew Bogyo

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Human DJ-1 is a highly conserved and yet functionally enigmatic protein associated with a heritable form of Parkinson’s disease. It has been suggested to be a redox-dependent regulatory scaffold, binding to proteins to modulate their function. Here we present the X-ray crystal structure of the Toxoplasma orthologue Toxoplasma gondii DJ-1 (TgDJ-1) at 2.1-Å resolution and show that it directly associates with calcium-dependent protein kinase 1 (CDPK1). The TgDJ-1 structure identifies an orthologously conserved arginine dyad that acts as a phospho-gatekeeper motif to control complex formation. We determined that the binding of TgDJ-1 to CDPK1 is sensitive to oxidation and calcium, and that this interaction potentiates CDPK1 kinase activity. Finally, we show that genetic deletion of TgDJ-1 results in upregulation of CDPK1 expression and that disruption of the CDPK1/TgDJ-1 complex in vivo prevents normal exocytosis of parasite virulence-associated organelles called micronemes. Overall, our data suggest that TgDJ-1 functions as a noncanonical kinase-regulatory scaffold that integrates multiple intracellular signals to tune microneme exocytosis in T. gondii.IMPORTANCE Apicomplexan parasites such as Toxoplasma and Plasmodium are obligate intracellular parasites that require the protective environment of a host cell in order to replicate and survive within a host organism. These parasites secrete effector proteins from specialized apical organelles to select and invade a chosen host cell. The secretion of these organelles is a tightly regulated process coordinated by endogenous small molecules and calcium-dependent protein kinases. We previously identified the Toxoplasma orthologue of the highly conserved protein DJ-1 as a regulator of microneme secretion, but the molecular basis for this was not known. We have now identified the molecular mechanism for how TgDJ-1 regulates microneme secretion. TgDJ-1 interacts with the kinase responsible for the secretion of these organelles (calcium-dependent kinase 1) and synergizes with calcium to potentiate kinase activity. This interaction is direct, phosphodependent, and necessary for the normal secretion of these important organelles.

Original languageEnglish (US)
Article numbere02189-16
JournalmBio
Volume8
Issue number1
DOIs
StatePublished - Jan 1 2017

Fingerprint

Toxoplasma
Organelles
Phosphotransferases
Parasites
Exocytosis
Calcium
calcium-dependent protein kinase
Plasmodium
Oxidation-Reduction
Parkinson Disease
Virulence
Arginine
Carrier Proteins
Proteins
Up-Regulation
X-Rays

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Child, M. A., Garland, M., Foe, I., Madzelan, P., Treeck, M., Van Der Linden, W. A., ... Bogyo, M. (2017). Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1. mBio, 8(1), [e02189-16]. https://doi.org/10.1128/mBio.02189-16

Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1. / Child, Matthew A.; Garland, Megan; Foe, Ian; Madzelan, Peter; Treeck, Moritz; Van Der Linden, Wouter A.; Bender, Kristina Oresic; Weerapana, Eranthie; Wilson, Mark A.; Boothroyd, John C.; Reese, Michael L.; Bogyo, Matthew.

In: mBio, Vol. 8, No. 1, e02189-16, 01.01.2017.

Research output: Contribution to journalArticle

Child, MA, Garland, M, Foe, I, Madzelan, P, Treeck, M, Van Der Linden, WA, Bender, KO, Weerapana, E, Wilson, MA, Boothroyd, JC, Reese, ML & Bogyo, M 2017, 'Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1', mBio, vol. 8, no. 1, e02189-16. https://doi.org/10.1128/mBio.02189-16
Child MA, Garland M, Foe I, Madzelan P, Treeck M, Van Der Linden WA et al. Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1. mBio. 2017 Jan 1;8(1). e02189-16. https://doi.org/10.1128/mBio.02189-16
Child, Matthew A. ; Garland, Megan ; Foe, Ian ; Madzelan, Peter ; Treeck, Moritz ; Van Der Linden, Wouter A. ; Bender, Kristina Oresic ; Weerapana, Eranthie ; Wilson, Mark A. ; Boothroyd, John C. ; Reese, Michael L. ; Bogyo, Matthew. / Toxoplasma DJ-1 regulates organelle secretion by a direct interaction with calcium-dependent protein kinase 1. In: mBio. 2017 ; Vol. 8, No. 1.
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