Proteomic analysis of the ventral disc of Giardia lamblia

Lourenço, Daniela; Andrade, Iamara da Silva; Terra, Letícia Labati; Guimarães, Patricia Ramos; Zingali, Russolina B.; Souza, Wanderley de

doi:10.1186/1756-0500-5-41

Cited by 16 publications

(7 citation statements)

References 25 publications

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“…A striking feature of Giardia encystation is disassembly of the adhesive disk [16] . We detected 22 of 38 previously published adhesive disk candidates [64] , [65] , 3 of which (GL50803_101291, 103676 and 13584) were at most 2-fold more abundant at 4hpie ( Table S2 ). This is consistent with previous reports of 2D PAGE analyses showing little change in adhesive disk protein abundance during encystation [16] .…”

Section: Resultsmentioning

confidence: 90%

The Proteome Landscape of Giardia lamblia Encystation

2013

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Giardia lamblia is an intestinal protozoan parasite required to survive in the environment in order to be transmitted to a new host. To ensure parasite survival, flagellated trophozoites colonizing the small intestine differentiate into non-motile environmentally-resistant cysts which are then shed in the environment. This cell differentiation process called encystation is characterized by significant morphological remodeling which includes secretion of large amounts of cyst wall material. Although much is known about the transcriptional regulation of encystation and the synthesis and trafficking of cyst wall material, the investigation of global changes in protein content and abundance during G. lamblia encystation is still unaddressed.In this study, we report on the quantitative analysis of the G. lamblia proteome during encystation using tandem mass spectrometry. Quantification of more than 1000 proteins revealed major changes in protein abundance in early, mid and late encystation, notably in constitutive secretory protein trafficking. Early stages of encystation were marked by a striking decrease of endoplasmic reticulum-targeted variant-specific surface proteins and significant increases in cytoskeleton regulatory components, NEK protein kinases and proteins involved in protein folding and glycolysis. This was in stark contrast to cells in the later stages of encystation which presented a surprisingly similar proteome composition to non-encysting trophozoites. Altogether these data constitute the first quantitative atlas of the Giardia proteome covering the whole process of encystation and point towards an important role for post-transcriptional control of gene expression in Giardia differentiation. Furthermore, our data provide a valuable resource for the community-based annotation effort of the G. lamblia genome, where almost 70% of all predicted gene models remains “hypothetical”.

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Section: Resultsmentioning

confidence: 90%

The Proteome Landscape of Giardia lamblia Encystation

2013

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“…These improvements for in vitro growth occurred within 2 weeks in 106-MtzR, while fluctuations in growth rate requiring higher seeding volumes at passage were observed for 713-MtzR for up to 4 weeks (P8). As both WB-MtzR and 713-MtzR have reduced abundance of beta-giardin and SALP-1 (4410) proteins of the ventral disc [ 58 ], it is possible both lines have lower attachment ( Supplementary Fig. S8 ) due to changes in the structure of the ventral disc and, therefore, lower confluence and slower division.…”

Section: Resultsmentioning

confidence: 99%

Differential protein expression and post-translational modifications in metronidazole-resistant Giardia duodenalis

et al. 2018

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BackgroundMetronidazole (Mtz) is the frontline drug treatment for multiple anaerobic pathogens, including the gastrointestinal protist, Giardia duodenalis. However, treatment failure is common and linked to in vivo drug resistance. In Giardia, in vitro drug-resistant lines allow controlled experimental interrogation of resistance mechanisms in isogenic cultures. However, resistance-associated changes are inconsistent between lines, phenotypic data are incomplete, and resistance is rarely genetically fixed, highlighted by reversion to sensitivity after drug selection ceases or via passage through the life cycle. Comprehensive quantitative approaches are required to resolve isolate variability, fully define Mtz resistance phenotypes, and explore the role of post-translational modifications therein.FindingsWe performed quantitative proteomics to describe differentially expressed proteins in 3 seminal Mtz-resistant lines compared to their isogenic, Mtz-susceptible, parental line. We also probed changes in post-translational modifications including protein acetylation, methylation, ubiquitination, and phosphorylation via immunoblotting. We quantified more than 1,000 proteins in each genotype, recording substantial genotypic variation in differentially expressed proteins between isotypes. Our data confirm substantial changes in the antioxidant network, glycolysis, and electron transport and indicate links between protein acetylation and Mtz resistance, including cross-resistance to deacetylase inhibitor trichostatin A in Mtz-resistant lines. Finally, we performed the first controlled, longitudinal study of Mtz resistance stability, monitoring lines after cessation of drug selection, revealing isolate-dependent phenotypic plasticity.ConclusionsOur data demonstrate understanding that Mtz resistance must be broadened to post-transcriptional and post-translational responses and that Mtz resistance is polygenic, driven by isolate-dependent variation, and is correlated with changes in protein acetylation networks.

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“…Palm et al [40] carried out a proteomic analysis of the cytoskeleton preparation and reported the presence of a family of giardins (α-1, β, γ, and δ) and two isoforms of tubulin and a new protein, SALP-1, which is homologous to proteins that participate in the aggregation of striate fibers. Subsequently, a proteomic analysis was carried out by Lourenço et al [41] using a cell fractionation approach. They obtained a highly enriched disk fraction that by SDS-PAGE showed the presence of five predominant bands, ranging from 25 to 58 kDa, as well as some light bands with higher molecular weight.…”

Section: Compositionmentioning

confidence: 99%

“…In recent years, proteomic approaches combined with microscope localization technique were carried out and new disk-associated proteins were identified such as the NIMA-related kinases (Neks), ankyrin repeat domain-containing protein, median body protein, and fungal cell wall protein Mp1p. These proteins have specific sites involved in cell adhesion and TTHERM, a hypothetical protein associated with the microtubule formation in the ciliate Tetrahymena thermophila [26,41]. However, despite the effort that has been made to characterize these structures by several research groups, the specific function of each of these proteins is not yet fully understood.…”

Section: Compositionmentioning

confidence: 99%

The Cytoskeleton of Giardia intestinalis

Gadelha¹,

Benchimol²,

Souza³

2017

Current Topics in Giardiasis

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Giardia intestinalis is a pathogenic protozoan, which is the causative agent of giardiasis. The Giardia trophozoite presents a cytoskeleton formed by specialized microtubular structures such as the ventral disk, four pairs of flagella, the median body, and the funis that are involved in cell division and differentiation. Because trophozoite motility and adhesion to the host intestinal cells are important processes mediated by the parasite cytoskeleton, the fine regulation of these elements may be directly related to the mechanisms that underlie infection. The organization of Giardia cytoskeleton at the ultrastructural level has been analyzed by different classical microscopy methods, including negative stain and chemical fixation for scanning and transmission electron microscopy. In this chapter, we provide an overview of the G. intestinalis cytoskeleton, emphasizing its structural organization and proteins involved in the maintenance of the structures as well as their functional role. These structures have been recently analyzed in some detail using techniques such as electron microscopy tomography, cryoelectron microscopy, ultra-high resolution scanning electron microscopy (UHRSEM), and helium ion microscopy (HIM). In addition, genome survey and phylogenetic analysis as well as proteomic analysis have revealed the presence of several new and not yet well-characterized proteins.

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Proteomic analysis of the ventral disc of Giardia lamblia

Cited by 16 publications

References 25 publications

The Proteome Landscape of Giardia lamblia Encystation

The Proteome Landscape of Giardia lamblia Encystation

Differential protein expression and post-translational modifications in metronidazole-resistant Giardia duodenalis

The Cytoskeleton of Giardia intestinalis

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