Functional analyses of a putative, membrane-bound, peroxisomal import mechanism from the apicomplexan protozoan Toxoplasma gondii

Mbekeani, A.J.; Stanley, W.A.; Kalel, V.C.; Dohan, N.; Zalckvar, E.; Scheiner, L.; Schliebs, W.; Erdmann, R.; Pohl, E.; Denny, P.W.

Abstract

Peroxisomes are central to eukaryotic metabolism, including the oxidation of fatty acids—which subsequently provide an important source of metabolic energy—and in the biosynthesis of cholesterol and plasmalogens. However, the presence and nature of peroxisomes in the parasitic apicomplexan protozoa remains controversial. A survey of the available genomes revealed that genes encoding peroxisome biogenesis factors, so-called peroxins (Pex), are only present in a subset of these parasites, the coccidia. The basic principle of peroxisomal protein import is evolutionarily conserved, proteins harbouring a peroxisomal-targeting signal 1 (PTS1) interact in the cytosol with the shuttling receptor Pex5 and are then imported into the peroxisome via the membrane-bound protein complex formed by Pex13 and Pex14. Surprisingly, whilst Pex5 is clearly identifiable, Pex13 and, perhaps, Pex14 are apparently absent from the coccidian genomes. To investigate the functionality of the PTS1 import mechanism in these parasites, expression of Pex5 from the model coccidian Toxoplasma gondii was shown to rescue the import defect of Pex5-deleted Saccharomyces cerevisiae. In support of these data, green fluorescent protein (GFP) bearing the enhanced (e)PTS1 known to efficiently localise to peroxisomes in yeast, localised to peroxisome-like bodies when expressed in Toxoplasma. Furthermore, the PTS1-binding domain of Pex5 and a PTS1 ligand from the putatively peroxisome-localised Toxoplasma sterol carrier protein (SCP2) were shown to interact in vitro. Taken together, these data demonstrate that the Pex5–PTS1 interaction is functional in the coccidia and indicate that a nonconventional peroxisomal import mechanism may operate in the absence of Pex13 and Pex14.

Citation

Mbekeani, A., Stanley, W., Kalel, V., Dohan, N., Zalckvar, E., Scheiner, L., Schliebs, W., Erdmann, R., Pohl, E., & Denny, P. (2018). Functional analyses of a putative, membrane-bound, peroxisomal import mechanism from the apicomplexan protozoan Toxoplasma gondii. Genes, 9(9), Article 434. https://doi.org/10.3390/genes9090434

• Journal Article Type: Article
• Acceptance Date: Aug 21, 2018
• Online Publication Date: Aug 29, 2018
• Publication Date: Aug 29, 2018
• Deposit Date: Jun 25, 2018
• Publicly Available Date: Aug 29, 2018
• Journal: Genes
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 9
• Issue: 9
• Article Number: 434
• DOI: https://doi.org/10.3390/genes9090434
• Public URL: https://durham-repository.worktribe.com/output/1323581