The nuclear envelope serves as an intermediary between the ER and Golgi complex in the intracellular parasite Toxoplasma gondii.
Morphological examination of the highly polarized protozoan parasite Toxoplasma gondii suggests that secretory traffic in this organism progresses from the endoplasmic reticulum to the Golgi apparatus using the nuclear envelope as an intermediate compartment. While the endoplasmic reticulum is predominantly located near the basal end of the parasite, the Golgi is invariably adjacent to the apical end of the nucleus, and the space between the Golgi and nuclear envelope is filled with numerous coatomer-coated vesicles. Staining with antiserum raised against recombinant T. gondii beta-COP confirms its association with the apical juxtanuclear region. Perturbation of protein secretion using brefeldin A, microtubule inhibitors or dithiothreitol disrupts the Golgi, causing swelling of the nuclear envelope, particularly at its basal end. Prolonged drug treatment leads to gross distention of the endoplasmic reticulum, filling the basal end of the parasite. Cloning and sequencing of the T. gondii homolog of the chaperonin protein BiP identifies the carboxy-terminal amino acid sequence HDEL as this organism's endoplasmic reticulum-retention signal. Appending the HDEL motif to a recombinant secretory protein (a chimera between the parasite's major surface protein fusion, P30, and the Green Fluorescent Protein) causes this secretory reporter to be retained intracellularly. P30-GFP-HDEL fluorescence was most intense within the nuclear envelope, particularly at the apical end. These data support a model of secretion in which protein traffic from the endoplasmic reticulum to Golgi occurs via the apical end of the nuclear envelope.
Hager
K M
KM
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104-6018, USA.
Striepen
B
B
Tilney
L G
LG
Roos
D S
DS
D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D020126:Brefeldin A / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D020756:Coatomer Protein / Q000032:analysis / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D000091342:Endoplasmic Reticulum Chaperone BiP; D004847:Epithelial Cells / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000235:genetics; D006056:Golgi Apparatus / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006360:Heat-Shock Proteins; D006801:Humans; D007202:Indicators and Reagents / Q000493:pharmacokinetics; D008164:Luminescent Proteins / Q000493:pharmacokinetics; D008854:Microscopy, Electron; D018832:Molecular Chaperones / Q000235:genetics; D009685:Nuclear Envelope / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D021382:Protein Sorting Signals / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D015800:Protozoan Proteins / Q000235:genetics; D012867:Skin / Q000166:cytology; D013045:Species Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014122:Toxoplasma / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
D002352:Carrier Proteins; D020756:Coatomer Protein; D000091342:Endoplasmic Reticulum Chaperone BiP; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D007202:Indicators and Reagents; C060153:KAR2 protein, yeast; D008164:Luminescent Proteins; D018832:Molecular Chaperones; D021382:Protein Sorting Signals; D011500:Protein Synthesis Inhibitors; D015800:Protozoan Proteins; D013439:Sulfhydryl Reagents; D049452:Green Fluorescent Proteins; D020126:Brefeldin A; D004229:Dithiothreitol