Rab7 and actin cytoskeleton participate during mobilization of β1<i>EH</i>FNR in fibronectin‐stimulated <i>Entamoeba histolytica</i>trophozoites

Javier-Reyna, Rosario; Hernández‐Ramírez, Verónica Ivonne; González‐Robles, Arturo; Galván-Mendoza, Ivan J.; Osorio-Trujillo, Juan Carlos; Talamás-Rohana, Patricia

doi:10.1002/jemt.21056

Cited by 13 publications

(15 citation statements)

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“…To date, ∼20 surface-exposed proteins or protein families of E. histolytica have been described (1–4, 6–15, 17–23, 36) (5, 16). To identify the entire set of plasma membrane-associated proteins, the surface molecules of E. histolytica were labeled with biotin, purified, and analyzed by mass spectrometry.…”

Section: Discussionmentioning

confidence: 99%

“…To date, only about 20 proteins or protein families have been identified as exposed on the plasma membrane of the parasite. These proteins include EhADH112 and the cysteine peptidase EhCP112 (EhCP-B9), which form a 112 kDa adhesion protein (3, 4); the serine-rich E. histolytica protein (SREHP) (5); a calreticulin (6); an as-yet unidentified mannose binding lectin (7); transmembrane kinases, including phagosome-associated TMK96 (PATMK), TMK39 and TMK54 (8, 9); a family of Bsp-A-like molecules (10); a rhomboid protease (11); an EhRab7 molecule (12); an actinin-like protein (AAF20148) (13); the lysine (K) and glutamic acid (E) enriched proteins KERP-1 and KERP-2 (13); the cysteine peptidases EhCP-A2 and EhCP-A5 (14, 15); a peroxiredoxin (29 kDa thiol-dependent peroxidase) (16); the transcription factor URE3-BP, which localizes to the cytoplasm and inner surface of the plasma membrane (17); the ARIEL antigen (18, 19); a LIM protein (EhLimA) associated with lipid rafts in the plasma membrane (20); the M8 family surface metalloprotease (EhMSP-1) (21); alcohol dehydrogenase 3 (22); and syntaxin 1 and SNAP-25 (23). …”

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confidence: 99%

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The Cell Surface Proteome of Entamoeba histolytica

Biller

Matthiesen

Kühne

et al. 2014

Molecular & Cellular Proteomics

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Surface molecules are of major importance for host-parasite interactions. During Entamoeba histolytica infections, these interactions are predicted to be of prime importance for tissue invasion, induction of colitis and liver abscess formation. To date, however, little is known about the molecules involved in these processes, with only about 20 proteins or protein families found exposed on the E. histolytica surface. We have therefore analyzed the complete surface proteome of E. histolytica. Using cell surface biotinylation and mass spectrometry, 693 putative surface-associated proteins were identified. In silico analysis predicted that ϳ26% of these proteins are membrane-associated, as they contain transmembrane domains and/or signal sequences, as well as sites of palmitoylation, myristoylation, or prenylation. An additional 25% of the identified proteins likely represent nonclassical secreted proteins. Surprisingly, no membraneassociation sites could be predicted for the remaining 49% of the identified proteins. To verify surface localization, 23 proteins were randomly selected and analyzed by immunofluorescence microscopy. Of these 23 proteins, 20 (87%) showed definite surface localization. These findings indicate that a far greater number of E. histolytica proteins than previously supposed are surface-associated, a phenomenon that may be based on the high membrane turnover of E. histolytica. Molecular & Cellular Proteomics 13: 10.1074/mcp.M113.031393, 132-144, 2014.The intestinal protozoan Entamoeba histolytica is an important human parasite. Its life cycle is relatively simple, consisting of infectious cysts that can survive outside the host and vegetative trophozoites that proliferate in the human gut. After infection, E. histolytica trophozoites are normally present in the intestine where they asymptomatically persist for months in the lumen. E. histolytica can become a pathogen by penetrating the intestinal mucosa and inducing colitis, or by disseminating to other organs, most commonly to the liver, where it induces abscess formation.The factors that determine the clinical outcomes of E. histolytica infections have not been well defined. Decisive factors may include genetic aspects of the host and/or parasite, the type of immune response mounted by the host, the presence of concomitant infections, and host diet. E. histolytica surface proteins are regarded to be of prime importance for hostparasite interactions. Members of the galactose/N-acetyl D-galactosamine-inhibitable (Gal/GalNAc) lectin family exposed on the surface of the parasite are considered important for adherence to target cells (1, 2), with adherence necessary for killing and/or phagocytosis. In addition to their involvement in adhesion and phagocytosis, the surface molecules of E. histolytica are exposed to the host's immune system. To date, only about 20 proteins or protein families have been identified as exposed on the plasma membrane of the parasite. These proteins include EhADH112 and the cysteine peptidase EhCP112 (EhCP-B9), which fo...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

The Cell Surface Proteome of Entamoeba histolytica

Biller

Matthiesen

Kühne

et al. 2014

Molecular & Cellular Proteomics

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“…ECM components have been used to coat surfaces where trophozoites are grown, for example collagen and FN (Ramírez-Tapia et al, 2015; Talamás-Lara et al, 2015), and these interaction trophozoite-substrate leads to a response as the differential expression of proteins, protease secretion (Piña-Vázquez et al, 2012), and cytoskeletal rearrangements in which actin polymerization is actively involved (Sengupta et al, 2009; Javier-Reyna et al, 2012; Starke et al, 2014; Aguilar-Rojas et al, 2016). EMC components (such as FN) have been used to generate micropatterns in order to stimulate eukaryotic cells to migrate, produce abundant and different membrane protrusions (Alamdari et al, 2013; Starke et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…During intestinal infection, trophozoites disrupt, and degrade intestinal mucosa barrier, and can also trogocyte mucosal epithelial cells and promote cell death by multiple cytotoxic mechanisms (Ralston et al, 2014; Begum et al, 2015). In the course of invasion, trophozoites interact with extracellular matrix components (ECM) such as fibronectin (FN), laminin and collagen (Solaymani-Mohammadi and Petri, 2008), which in vitro induce actin cytoskeleton remodeling that is involved in adhesion, migration (Sengupta et al, 2009; Javier-Reyna et al, 2012) and motility (Aguilar-Rojas et al, 2016). …”

Section: Introductionmentioning

confidence: 99%

Influence of Micropatterned Grill Lines on Entamoeba histolytica Trophozoites Morphology and Migration

Sierra-López

Baylón-Pacheco

Espíritu-Gordillo

et al. 2018

Front. Cell. Infect. Microbiol.

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Entamoeba histolytica, the causal agent of human amoebiasis, has two morphologically different phases: a resistant cyst and a trophozoite responsible for the invasion of the host tissues such as the colonic mucosa and the intestinal epithelium. During in vitro migration, trophozoites usually produce protuberances such as pseudopods and rarely filopodia, structures that have been observed in the interaction of trophozoites with human colonic epithelial tissue. To study the different membrane projections produced by the trophozoites, including pseudopods, filopodia, uropods, blebs, and others, we designed an induction system using erythrocyte extract or fibronectin (FN) in micropatterned grill lines (each micro-line containing multiple micro-portions of FN or erythrocyte extract) on which the trophozoites were placed in culture for migration assays. Using light, confocal, and scanning electron microscopy, we established that E. histolytica trophozoites frequently produce short and long filopodia, large retractile uropods in the rear, pseudopods, blebs, and others structures, also showing continuous migration periods. The present study provides a simple migration method to induce trophozoites to generate abundant membrane protrusion structures that are rarely obtained in normal or induced cultures, such as long filopodia; this method will allow a–better understanding of the interactions of trophozoites with FN and cell debris. E. histolytica trophozoites motility plays an important role in invasive amoebiasis. It has been proposed that both physical forces and chemical signals are involved in the trophozoite motility and migration. However, the in vivo molecules that drive the chemotactic migration remain to be determined. We propose the present assay to study host molecules that guide chemotactic behavior because the method is highly reproducible, and a live image of cell movement and migration can be quantified.

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“…Surface localization implies vesicle traffi cking and recycling, independent from de novo protein synthesis [ 38 ]. The actin cytoskeleton and Rab7, a protein involved in the regulation of vesicular traffi cking, participate in the relocalization of β1FNR upon FN contact [ 70 ].…”

Section: The E Histolytica Fibronectin Receptor β1fnrmentioning

confidence: 99%

Cell-Surface Molecules as Virulence Determinants in Entamoeba histolytica

Faust

Guillén

2014

Amebiasis

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Virulence of the human parasite Entamoeba histolytica is related to adherence to, killing, and phagocytosis of host cells, and contact between trophozoites and host cells is necessary for amoebic effectors to act. Virulence also relies on parasite motility in extracellular matrices and human tissues and on the capacity of the parasite to escape the host defense response and to trigger infl ammation. Amoebic surface components implicated in all these processes have been identifi ed as important virulence factors. This chapter presents current knowledge on the major surface molecules of E. histolytica critically involved in parasite interactions with healthy or apoptotic cells (Gal/GalNAc lectin, LPG/LPPG proteophosphoglycans, KERP1, CPADH, STIRP, proform of CP-A5, ROM1), in recognition of apoptotic cells (calreticulin, M17, C2PK, SREHP, TMK96), host cell phagocytosis (CPADH, SREHP, TMK96, ROM1), attachment to extracellular matrices (β1FNR, β2 integrinlike), and immune evasion or triggering the infl ammatory response (Gal/GalNAc lectin, KERP1, proform of CP-A5, LPG/LPPG).

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Rab7 and actin cytoskeleton participate during mobilization of β1EHFNR in fibronectin‐stimulated Entamoeba histolyticatrophozoites

Cited by 13 publications

References 45 publications

The Cell Surface Proteome of Entamoeba histolytica

The Cell Surface Proteome of Entamoeba histolytica

Influence of Micropatterned Grill Lines on Entamoeba histolytica Trophozoites Morphology and Migration

Cell-Surface Molecules as Virulence Determinants in Entamoeba histolytica

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