Identification of a Golgi apparatus protein complex important for the asexual erythrocytic cycle of the malaria parasite
            <i>Plasmodium falciparum</i>

Hallée, Stéphanie; Thériault, Catherine; Gagnon, Dominic; Kehrer, Jessica; Frischknecht, Friedrich; Mair, Gunnar R.; Richard, Dave

doi:10.1111/cmi.12843

Cited by 10 publications

(8 citation statements)

References 57 publications

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“…It is noteworthy that in the mature female gametocyte, the ER is most dense in the polar regions of the parasite. This coincides with the distribution of a closely associated organelle, the Golgi, which, in Plasmodium , takes the rudimentary form of dispersed unstacked cisternae and is recognizable as a smooth membraned vesicle cluster 22, 40–42 . In females, the Golgi is found in two distinct clusters at both polar ends, while in the mature male it appears in four smaller clusters, similar to the developing gametocytes that also have a more widespread distribution.…”

Section: Resultssupporting

confidence: 69%

Comparative 3D ultrastructure ofPlasmodium falciparumgametocytes

Evers

Roverts

Boshoven

et al. 2023

Preprint

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Despite the enormous significance of malaria parasites for global health, some basic features of their ultrastructure remain obscure. In this study, we apply high-resolution volumetric electron microscopy to examine and compare the ultrastructure of Plasmodium falciparum gametocytes of both genders and in different stages of development as well as the more intensively studied asexual blood stages revisiting previously described phenomena in 3D. In doing so, we challenge the widely accepted notion of a single mitochondrion by demonstrating the presence of multiple mitochondria in gametocytes. We also provide evidence for a gametocyte-specific cytostome variant. Furthermore, we generate, among other organelles, the first 3D reconstructions of endoplasmic reticulum (ER), Golgi apparatus, and exported structures in gametocytes. Assessing interconnectivity between organelles, we find frequent structural appositions between the nucleus, mitochondria, and apicoplast. We provide evidence that the ER is a promiscuous interactor with numerous organelles and the trilaminar membrane of the gametocyte. Public availability of these volumetric electron microscopy resources of wild-type asexual and sexual blood-stage malaria parasites will facilitate reinterrogation of this global dataset with different research questions and expertise. Taken together, we reconstruct the 3D ultrastructure of P. falciparum gametocytes in high detail and shed light on the unique organellar biology of these deadly parasites.

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Section: Resultssupporting

confidence: 69%

Comparative 3D ultrastructure ofPlasmodium falciparumgametocytes

Evers

Roverts

Boshoven

et al. 2023

Preprint

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“…26 ). For example, the system is now commonly used in the apicomplexan malaria parasite Plasmodium falciparum for the analysis of drug targets as well as essential genes and processes 30,[55][56][57][58] . A glmS ribozyme system has also been established for yeast and, very recently, for T. brucei and T. cruzi 31,59,60 .…”

Section: Discussionmentioning

confidence: 99%

Testing the CRISPR-Cas9 and glmS ribozyme systems in Leishmania tarentolae

Turra

Schneider

Liedgens

et al. 2021

Molecular and Biochemical Parasitology

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Leishmania parasites include important pathogens and model organisms and are even used for the production of recombinant proteins. However, functional genomics and the characterization of essential genes are often limited in Leishmania because of low-throughput technologies for gene disruption or tagging and the absence of components for RNA interference. Here, we tested the T7 RNA polymerase-dependent CRISPR-Cas9 system by Beneke et al. and the glmS ribozyme-based knockdown system in the model parasite Leishmania tarentolae. We successfully deleted two reference genes encoding the flagellar motility factor Pf16 and the salvage-pathway enzyme adenine phosphoribosyltransferase, resulting in immotile and drug-resistant parasites, respectively. In contrast, we were unable to disrupt the gene encoding the mitochondrial flavoprotein Erv. Cultivation of L. tarentolae in standard BHI medium resulted in a constitutive down-regulation of an episomal mCherry-glmS reporter by 40 to 60%. For inducible knock-downs, we evaluated the growth of L. tarentolae in alternative media and identified supplemented MEM, IMDM and McCoy's 5A medium as candidates. Cultivation in supplemented MEM allowed an inducible, glucosamine concentrationdependent down-regulation of the episomal mCherry-glmS reporter by more than 70%. However, chromosomal glmS-tagging of the genes encoding Pf16, adenine phosphoribosyltransferase or Erv did not reveal a knock-down phenotype. Our data demonstrate the suitability of the CRISPR-Cas9 system for the disruption and tagging of genes in L. tarentolae as well as the limitations of the glmS system, which was restricted to moderate efficiencies for episomal knock-downs and caused no detectable phenotype for chromosomal knock-downs.

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“…Our results suggest that E. histolytica and E. dispar may have a rudimentary Golgi apparatus being composed of stacked cisternae, dispersed after monensin treatment and colocalizing with the Golgi marker NBD 6 ceramide. This Golgi apparatus in amoeba should be more similar to other parasites as Plasmodium falciparum (Hallée et al, 2018) than to the classical composition from Golgi apparatus in higher eukaryotes. However, which proteins related with Golgi apparatus in amoeba could participate in the intracellular trafficking machinery and how organelle biogenesis occurs are still under discover.…”

Section: Discussionmentioning

confidence: 86%

Golgi apparatus components in Entamoeba histolytica and Entamoeba dispar after monensin treatment

Talamás‐Lara

Acosta-Virgen

Chávez‐Munguía

et al. 2021

Microscopy Res & Technique

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Highly dynamic ribosomes, glycogen granules, thinly fibrillar material, and multiple membrane-bound vesicles are embedded in the matrix-rich cytoplasm of Entamoeba spp. trophozoites. The absence of a Golgi apparatus in these amoebae has been commonly accepted. Here we challenge this observation by incubating Entamoeba histolytica and Entamoeba dispar with monensin, an ionophore that produces swelling of the Golgi apparatus. We observe changes in the trophozoites through standard transmission electron microscopy, cryofixation and cryosubstitution, and analyze the label and expression of known resident proteins of the cis-GM130 and trans-TGN38 Golgi network through confocal microscopy and Western blot assays. Cryosubstitution and standard methods using the treatment, preserved membranous lamellae resembling Golgi components. GM130 and TGN38 Golgi antigens were found by immunoelectron, immunoblot, and co-localization by confocal microscopy using the reagent NBD C6-ceramide. Our results indicate that previously undetected Golgi apparatus components are present in the cytoplasm of E. histolytica and E. dispar.

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Identification of a Golgi apparatus protein complex important for the asexual erythrocytic cycle of the malaria parasite Plasmodium falciparum

Cited by 10 publications

References 57 publications

Comparative 3D ultrastructure ofPlasmodium falciparumgametocytes

Comparative 3D ultrastructure ofPlasmodium falciparumgametocytes

Testing the CRISPR-Cas9 and glmS ribozyme systems in Leishmania tarentolae

Golgi apparatus components in Entamoeba histolytica and Entamoeba dispar after monensin treatment

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