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Targeting to rhoptry organelles of Toxoplasma gondii involves evolutionarily conserved mechanisms.

Nature Cell Biology volume 2, pages 449–456 (2000)Cite this article

Abstract

Intracellular parasites of the phylum Apicomplexa contain specialized rhoptry secretory organelles that have a crucial function in host-cell invasion and establishment of the parasitophorous vacuole. Here we show that localization of the Toxoplasma gondii rhoptry protein ROP2 is dependent on a YEQL sequence in the cytoplasmic tail that binds to µ-chain subunits of T. gondii and mammalian adaptors, and conforms to the YXXφ mammalian sorting motif. Chimaeric reporters, containing the transmembrane domains and cytoplasmic tails of the low-density lipoprotein receptor and of Lamp-1, are sorted to the Golgi or the trans-Golgi network (TGN), and partially to apical microneme organelles of the parasite, respectively. Targeting of these reporters is mediated by YXXφ- and NPXY-type signals. This is the first demonstration of tyrosine-dependent sorting in protozoan parasites, indicating that T. gondii proteins may be targeted to, and involved in biogenesis of, morphologically unique organelles through the use of evolutionarily conserved signals and machinery.

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Figure 1: Constructs used in this study.
Figure 2: The cytoplasmic YEQL motif is necessary for targeting of ROP2 to rhoptry organelles.
Figure 3: The SEYEQL sequence of ROP2 interacts with clathrin adaptor µ-chains in a yeast two-hybrid growth assay.
Figure 4: The GYQTI motif directs sorting of the B–Lamp chimaera in T. gondii.
Figure 5: B–Lamp partially co-localizes with micronemes in T. gondii.
Figure 6: Immunoelectron-microscopic localization of BAP chimaeras in T.gondii .
Figure 7: Tyrosine-dependent localization of B–LDLR constructs in T. gondii.
Figure 8: Characterization of HA-tagged ROP2(ΔYEQL).
Figure 9: Tyrosine-dependent protein sorting in T. gondii.

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Acknowledgements

This work was supported by Public Health Service Grant ROI AI30060 from the National Institutes of Health and a Scholar Award in Molecular Parasitology from the Burroughs Wellcome Fund to K.A.J., by National Institutes of Health training grant 5T32AI-07404 to H.M.N., by a South African Foundation for Research Development postdoctoral fellowship to H.C.H, and by an individual National Research Service Award 5F32AI-10044 to H.M.N. We thank N. Andrews for critical reading of the manuscript.

Correspondence and requests for materials should be addressed to K.A.J.

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  1. Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, 06520-8022, Connecticut, USA

    Heinrich C. Hoppe, Huân M. Ngô, Mei Yang & Keith A. Joiner

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Correspondence to Keith A. Joiner.

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Hoppe, H., Ngô, H., Yang, M. et al. Targeting to rhoptry organelles of Toxoplasma gondii involves evolutionarily conserved mechanisms.. Nat Cell Biol 2, 449–456 (2000). https://doi.org/10.1038/35017090

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