Cyst-Wall-Protein-1 is fundamental for Golgi-like organelle neogenesis and cyst-wall biosynthesis in Giardia lamblia

Ebneter, Jacqueline A.; Heusser, Sally D.; Schraner, Elisabeth M.; Hehl, Adrian B.; Faso, Carmen

doi:10.1038/ncomms13859

Cited by 35 publications

(45 citation statements)

References 54 publications

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“…CWP1 (Ebneter et al, 2016) was identified as differentially expressed by only one of the two methods of analysis we used, and was thus not included in the 475 differentially expressed genes (Materials and Methods). Through partitioning analysis and analysis of differential gene expression in the in vivo foci, we also identified six new cyst-specific proteins (Figure 5D and Supplemental Tables 7,8).…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic Profiling of High-Density Giardia Foci Encysting in the Murine Proximal Intestine

Pham

Nosala

Scott

et al. 2017

Front. Cell. Infect. Microbiol.

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Giardia is a highly prevalent, understudied protistan parasite causing significant diarrheal disease worldwide. Its life cycle consists of two stages: infectious cysts ingested from contaminated food or water sources, and motile trophozoites that colonize and attach to the gut epithelium, later encysting to form new cysts that are excreted into the environment. Current understanding of parasite physiology in the host is largely inferred from transcriptomic studies using Giardia grown axenically or in co-culture with mammalian cell lines. The dearth of information about the diversity of host-parasite interactions occurring within distinct regions of the gastrointestinal tract has been exacerbated by a lack of methods to directly and non-invasively interrogate disease progression and parasite physiology in live animal hosts. By visualizing Giardia infections in the mouse gastrointestinal tract using bioluminescent imaging (BLI) of tagged parasites, we recently showed that parasites colonize the gut in high-density foci. Encystation is initiated in these foci throughout the entire course of infection, yet how the physiology of parasites within high-density foci in the host gut differs from that of cells in laboratory culture is unclear. Here we use BLI to precisely select parasite samples from high-density foci in the proximal intestine to interrogate in vivo Giardia gene expression in the host. Relative to axenic culture, we noted significantly higher expression (>10-fold) of oxidative stress, membrane transporter, and metabolic and structural genes associated with encystation in the high-density foci. These differences in gene expression within parasite foci in the host may reflect physiological changes associated with high-density growth in localized regions of the gut. We also identified and verified six novel cyst-specific proteins, including new components of the cyst wall that were highly expressed in these foci. Our in vivo transcriptome data support an emerging view that parasites encyst early in localized regions in the gut, possibly as a consequence of nutrient limitation, and also impact local metabolism and physiology.

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Section: Discussionmentioning

confidence: 99%

Transcriptomic Profiling of High-Density Giardia Foci Encysting in the Murine Proximal Intestine

Pham

Nosala

Scott

et al. 2017

Front. Cell. Infect. Microbiol.

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“…Given that CCVs have not been detected in Giardia , this begs the question of the functional role of a NECAP1 cardiolipin-binding orthologue in G. lamblia which was found to interact with G. lamblia AP2 subunits and Gl FYVE. Recent developments in gene knock-out [63] and CRISPR-Cas9-based knock-down [64, 65] methodologies tailored to G. lamblia will be instrumental towards a full functional characterization of Gl NECAP1’s function(s)…”

Section: Discussionmentioning

confidence: 99%

Phosphoinositide-binding proteins mark, shape and functionally modulate highly-diverged endocytic compartments in the parasitic protistGiardia lamblia

Černíková

Faso

Hehl

2019

Preprint

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Phosphorylated derivatives of phosphatidylinositol (PIPs), are key membrane lipid residues involved in clathrin-mediated endocytosis (CME). CME relies on PI(4,5)P2 to mark endocytic sites at the plasma membrane (PM) associated to clathrin-coated vesicle (CCV) formation. The highly diverged parasitic protist Giardia lamblia presents disordered and static clathrin assemblies at PM invaginations, contacting specialized endocytic organelles called peripheral vacuoles (PVs). The role for clathrin assemblies in fluid phase uptake and their link to internal membranes via PIP-binding adaptors is unknown.Here we provide evidence for a robust link between clathrin assemblies and fluid-phase uptake in G. lamblia mediated by proteins carrying predicted PX, FYVE and NECAP1 PIP-binding modules. We show that chemical and genetic perturbation of PIP-residue binding and turnover elicits novel uptake and organelle-morphology phenotypes. A combination of co-immunoprecipitation and in silico annotation techniques expands the initial PIP-binding network with addition of new members. Our data indicate that, despite the partial conservation of lipid markers and protein cohorts known to play important roles in dynamic endocytic events in well-characterized model systems, the Giardia lineage presents a strikingly divergent clathrin-centered network. This includes several PIP-binding modules, often associated to domains of currently unknown function that shape and modulate fluid-phase uptake at PVs.

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“…Given that CCVs have not been detected in Giardia, this begs the question of the functional role of a NECAP1 cardiolipin-binding orthologue in G. lamblia, which was found to interact with G. lamblia AP2 subunits and GlFYVE. Recent developments in gene knock-out [67] and CRISPR-Cas9-based knock-down [68,69] methodologies tailored to G. lamblia will be instrumental towards a full functional characterization of GlNE-CAP1's function(s).…”

Section: An Interactome-based Model For Pip-binding Proteins and Clatmentioning

confidence: 99%

Phosphoinositide-binding proteins mark, shape and functionally modulate highly-diverged endocytic compartments in the parasitic protist Giardia lamblia

2020

Self Cite

View full text Add to dashboard Cite

Phosphorylated derivatives of phosphatidylinositol (PIPs) are key membrane lipid residues involved in clathrin-mediated endocytosis (CME). CME relies on PIP species PI(4,5)P 2 to mark endocytic sites at the plasma membrane (PM) associated to clathrin-coated vesicle (CCV) formation. The highly diverged parasitic protist Giardia lamblia presents disordered and static clathrin assemblies at PM invaginations, contacting specialized endocytic organelles called peripheral vacuoles (PVs). The role for clathrin assemblies in fluid phase uptake and their link to internal membranes via PIP-binding adaptors is unknown. Here we provide evidence for a robust link between clathrin assemblies and fluid-phase uptake in G. lamblia mediated by proteins carrying predicted PX, FYVE and NECAP1 PIP-binding modules. We show that chemical and genetic perturbation of PIP-residue binding and turnover elicits novel uptake and organelle-morphology phenotypes. A combination of co-immunoprecipitation and in silico analysis techniques expands the initial PIP-binding network with addition of new members. Our data indicate that, despite the partial conservation of lipid markers and protein cohorts known to play important roles in dynamic endocytic events in well-characterized model systems, the Giardia lineage presents a strikingly divergent clathrin-centered network. This includes several PIP-binding modules, often associated to domains of currently unknown function that shape and modulate fluid-phase uptake at PVs.

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Cyst-Wall-Protein-1 is fundamental for Golgi-like organelle neogenesis and cyst-wall biosynthesis in Giardia lamblia

Cited by 35 publications

References 54 publications

Transcriptomic Profiling of High-Density Giardia Foci Encysting in the Murine Proximal Intestine

Transcriptomic Profiling of High-Density Giardia Foci Encysting in the Murine Proximal Intestine

Phosphoinositide-binding proteins mark, shape and functionally modulate highly-diverged endocytic compartments in the parasitic protistGiardia lamblia

Phosphoinositide-binding proteins mark, shape and functionally modulate highly-diverged endocytic compartments in the parasitic protist Giardia lamblia

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