A role for adaptor protein complex 1 in protein targeting to rhoptry organelles in Plasmodium falciparum

• Published in: Biochimica et biophysica acta 2015-03, Vol.1853 (3), p.699-710
• Main Authors: Kaderi Kibria, K.M., Rawat, Khushboo, Klinger, Christen M., Datta, Gaurav, Panchal, Manoj, Singh, Shailja, Iyer, Gayatri R., Kaur, Inderjeet, Sharma, Veena, Dacks, Joel B., Mohmmed, Asif, Malhotra, Pawan
• Format: Article
• Language: English
• Subjects: Adaptor Protein Complex 1 - physiology; Adaptor protein complex-1 (AP-1); Cells, Cultured; Clathrin; Erythrocytes - parasitology; ER–Golgi network; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Membrane Proteins - metabolism; Organelles - metabolism; Organisms, Genetically Modified; Plasmodium falciparum; Plasmodium falciparum - genetics; Plasmodium falciparum - growth & development; Plasmodium falciparum - metabolism; Protein Transport - genetics; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Trafficking; Vesicular trafficking
• ISSN: 0167-4889; 0006-3002; 1879-2596
• DOI: 10.1016/j.bbamcr.2014.12.030

The human malaria parasite Plasmodium falciparum possesses sophisticated systems of protein secretion to modulate host cell invasion and remodeling. In the present study, we provide insights into the function of the AP-1 complex in P. falciparum. We utilized GFP fusion constructs for live cell imaging, as well as fixed parasites in immunofluorescence analysis, to study adaptor protein mu1 (Pfμ1) mediated protein trafficking in P. falciparum. In trophozoites Pfμ1 showed similar dynamic localization to that of several Golgi/ER markers, indicating Golgi/ER localization. Treatment of transgenic parasites with Brefeldin A altered the localization of Golgi-associated Pfμ1, supporting the localization studies. Co-localization studies showed considerable overlap of Pfμ1 with the resident rhoptry proteins, rhoptry associated protein 1 (RAP1) and Cytoadherence linked asexual gene 3.1 (Clag3.1) in schizont stage. Immunoprecipitation experiments with Pfμ1 and PfRAP1 revealed an interaction, which may be mediated through an intermediate transmembrane cargo receptor. A specific role for Pfμ1 in trafficking was suggested by treatment with AlF4, which resulted in a shift to a predominantly ER-associated compartment and consequent decrease in co-localization with the Golgi marker GRASP. Together, these results suggest a role for the AP-1 complex in rhoptry protein trafficking in P. falciparum. •In Plasmodium falciparum four complexes (AP 1–4) have been identified by in-silico studies•Transgenic line was generated expressing Pfμ1–GFP fusion protein•Pfμ1 is associated with the ER–Golgi region and with rhoptries in developing merozoites.•Proteomic analysis suggests the role of PfAP-1 complex in protein targeting to rhoptry.