Spatial dissection of the cis‐ and trans‐Golgi compartments in the malaria parasite Plasmodium falciparum

Abstract

SummaryThe Golgi apparatus forms the heart of the secretory pathway in eukaryotic cells where proteins are modified, processed and sorted. The transport of proteins from the endoplasmic reticulum (ER) to the cis-side of the Golgi complex takes place at specialized ER subdomains known as transitional ER (tER). We used the Plasmodium falciparum orthologue of Sec13p to analyse tER organization. We show that the distribution of PfSec13p is restricted to defined areas of the ER membrane. These foci are juxtaposed to the Golgi apparatus and might represent tER sites. To further analyse cis-to trans-Golgi architecture, we generated a double transfectant parasite line that expresses the Golgi marker Golgi reassembly stacking protein (GRASP) as a green fluorescent protein fusion and the trans-Golgi marker Rab6 as a DsRed fusion protein. Our data demonstrate that Golgi multiplication is closely linked to tER multiplication, and that parasite maturation is accompanied by the spatial separation of the cis-and trans-face of this organelle.

Organellar proteomics reveals hundreds of novel nuclear proteins in the malaria parasite Plasmodium falciparum

“…Primary antibody dilutions were: rat mAb anti-HA 3F10 (Roche Diagnostics) 1:100; mouse mAb anti-Ty BB2 1:2,000 (kind gift from Keith Gull); mouse mAb anti-GAPDH 1:500 [128]; rabbit anti-PfBIP 1:500 [132,133] (kind gift from Tim Gilberger); and mouse mAb anti-fibrillarin 1:500 [71] (kind gift from Michael Terns, University of Georgia). Secondary antibody dilutions were: Alexa-Fluor ® 568-conjugated anti-rat IgG (Molecular Probes) 1:500; Alexa-Fluor ® 488-conjugated anti-rabbit IgG (Molecular Probes) 1:500; and FITC-conjugated anti-mouse IgG (Kirkegaard Perry Laboratories) 1:250.…”

Plasmodium falciparumATG8 implicated in both autophagy and apicoplast formation

“…The complete coding sequence (CDS) of PfATG8 (375 bp) and PfRAB7 (621 bp) were PCR amplified from cDNA of 3D7 P. falciparum (Fig. S1) and cloned into the AvrII/XhoI site of the P. falciparum expression vector pARL-GFP 44 to generate GFP-PfATG8 and GFP-PfRAB7. The primers used were: 5´-TACCCTAGGA TGCCATCGCT TAAAG-3´ (PfATG8 sense); 5´-TAGCTCGAGT TATCCTAGAC AACTCTC-3´ (PfATG8 antisense) and 5´-TTACCTAGGA TGTCAAATAA AAAAAGAACC-3´ (PfRAB7 sense); 5´-CATCTCGAGT TAACAACAAC GACTTTTG-3´ (PfRAB7 antisense).…”

Neogenesis and maturation of transient Golgi-like cisternae in a simple eukaryote

“…In unicellular organisms, Golgi organization is less complex, ranging from unlinked stacks associated with ER exit sites in Pichia pastoris (Bevis et al, 2002;Rossanese et al, 1999) and several protozoa (Benchimol et al, 2001;Hager et al, 1999;Struck et al, 2008), to individual, functionally distinct Golgi cisternae without stacked organization in Saccharomyces cerevisiae (Rossanese et al, 1999). Microsporidia, intracellular fungal parasites of mammals, have highly reduced trafficking systems, and lack a conventional Golgi (Beznoussenko et al, 2007).…”