Entamoeba invadens: Identification of a SERCA protein and effect of SERCA inhibitors on encystation

Martínez-Higuera, Aarón; Herrera-Martínez, Mayra; Chávez‐Munguía, Bibiana; Valle-Solis, Martha I.; Muñiz-Lino, None Marcos Agustín; Cázares-Apátiga, Javier; Rodríguez, Mario A.

doi:10.1016/j.micpath.2015.08.016

Cited by 12 publications

(9 citation statements)

References 35 publications

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“…Still, parasite SERCA/ATP6 seems to hold potential as a therapeutic target, because thapsigargin (Table 3 ), a plant sesquiterpene lactone and general SERCA inhibitor, killed cultured chloroquine resistant and sensitive P. falciparum parasites with IC 50 of 246 and 298 nM (del Pilar Crespo et al, 2008 ; Abiodun et al, 2013 ). Thapsigargin further inhibited growth of T. gondii, Trypanosoma spp., L. donovani, E. invadens , and Neospora canium with EC 50 -values in the range of 0.5–39 μM (Kim et al, 2002 ; Mishina et al, 2007 ; Martínez-Higuera et al, 2015 ). Yeast expressed T. gondii SERCA was fully inhibited by thapsigargin at 1 μM (Nagamune et al, 2007 ).…”

Section: Targeting Transport Processes Of Parasites At Different Levementioning

confidence: 99%

Targeting Channels and Transporters in Protozoan Parasite Infections

2018

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Infectious diseases caused by pathogenic protozoa are among the most significant causes of death in humans. Therapeutic options are scarce and massively challenged by the emergence of resistant parasite strains. Many of the current anti-parasite drugs target soluble enzymes, generate unspecific oxidative stress, or act by an unresolved mechanism within the parasite. In recent years, collections of drug-like compounds derived from large-scale phenotypic screenings, such as the malaria or pathogen box, have been made available to researchers free of charge boosting the identification of novel promising targets. Remarkably, several of the compound hits have been found to inhibit membrane proteins at the periphery of the parasites, i.e., channels and transporters for ions and metabolites. In this review, we will focus on the progress made on targeting channels and transporters at different levels and the potential for use against infections with apicomplexan parasites mainly Plasmodium spp. (malaria) and Toxoplasma gondii (toxoplasmosis), with kinetoplastids Trypanosoma brucei (sleeping sickness), Trypanosoma cruzi (Chagas disease), and Leishmania ssp. (leishmaniasis), and the amoeba Entamoeba histolytica (amoebiasis).

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Section: Targeting Transport Processes Of Parasites At Different Levementioning

confidence: 99%

Targeting Channels and Transporters in Protozoan Parasite Infections

2018

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“…Most developmental studies have been done in a closely related reptilian parasite, E. invadens , which can be encysted in vitro using glucose depletion and osmotic stress (Avron et al, 1986 ) and excysted from cysts to trophozoites using media supplemented with glucose, bile salt, sodium bicarbonate and serum (Mitra and Krishna Murti, 1978 ). Using this model of Entamoeba development, a number of triggers of encystation including catecholamine, gal-terminated ligands, cyclic AMP (cAMP), cholesteryl sulfate, NAD + , Ca 2+ signaling, and phospholipase-D (PLD) have been identified (Chayen et al, 1985 ; Cho and Eichinger, 1998 ; Eichinger, 2001 ; Makioka et al, 2001 ; Coppi et al, 2002 ; Frederick and Eichinger, 2004 ; Ehrenkaufer et al, 2013 ; Martinez-Higuera et al, 2015 ; Mi-ichi et al, 2015 ; Manna et al, 2018 ; Manna and Singh, 2019 ). Furthermore, a number of molecules, e.g., galactose, N-acetylglucosamine, and short chain fatty acids, have been shown to inhibit encystation (Coppi and Eichinger, 1999 ; Byers et al, 2005 ).…”

Section: Introductionmentioning

confidence: 99%

The NAD+ Responsive Transcription Factor ERM-BP Functions Downstream of Cellular Aggregation and Is an Early Regulator of Development and Heat Shock Response in Entamoeba

Manna

Lozano-Amado

Ehrenkaufer

et al. 2020

Front. Cell. Infect. Microbiol.

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Entamoeba histolytica is a protozoan parasite and a major cause of dysentery and diarrheal disease in developing countries. Disease transmission from one host to another occurs via cysts which can survive in environmental extremes and are transmitted through contaminated food and water. Recent studies in our lab identified a novel transcription factor, Encystation Regulatory Motif-Binding Protein (ERM-BP), which is responsive to NAD + and has an important role in encystation. The key residues important for ERM-BP function were demonstrated in vitro using recombinant protein. In this study we demonstrate the in vivo functional consequences of mutations in key domains and their impact on Entamoeba encystation. Our results show that mutations in the DNA binding domain (ERM-BP-DBM) and in the nicotinamidase domain (ERM-BP-C198A) lead to protein mis-localization in both trophozoites and cysts and significantly reduce encystation efficiency. Additionally, we showed that silencing of ERM-BP significantly decreased the size and number of multi-nucleated giant cells (MGC) that form during encystation, indicating that ERM-BP functions upstream of the cellular aggregation that precedes stage conversion. Dissection of epistatic interactions between ERM-BP and a second encystation-related transcription factor, NF-Y revealed that ERM-BP is upstream of NF-Y in controlling the developmental cascade and appears to be one of the earliest regulators of development identified to date in Entamoeba. We also demonstrated that ERM-BP is upregulated during heat stress in Entamoeba, another condition which increases intracellular NAD + levels and that overexpression of ERM-BP makes E. histolytica and E. invadens parasites more resistant to heat stress. Overexpression of ERM-BP in E. histolytica also induced the formation of cyst-like quadrinucleated cells and formation of MGCs. Overall, our work has identified an important role of ERM-BP in Entamoeba stress response and links an NAD +-responsive transcription factor to both development and heat shock response. Characterization of stress and developmental cascades are important avenues to investigate for Entamoeba, an important human parasitic pathogen.

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“…Our previous studies showed that E. histolytica contains at least five Ca 2+ -ATPases: three related to PMCAs, one to Sarco/Endoplasmic reticulum Ca 2+ -ATPases (SERCA), and another to Secretory Pathway Ca 2+ -ATPases (SPCA) (Martinez-Higuera et al, 2013 ; Rodríguez et al, 2018 ). Indeed, we detected the SPCA-related pump in the Golgi apparatus of E. histolytica (Rodríguez et al, 2018 ) and the SERCA-related pump in the endoplasmic reticulum of E. histolytica and E. invadens (Martinez-Higuera et al, 2013 ; Martínez-Higuera et al, 2015 ). In addition, we demonstrated that specific inhibitors of SERCA affected the encystation of E. invadens (Martínez-Higuera et al, 2015 ), suggesting that calcium flux through SERCA is involved in the development of Entamoeba sp.…”

Section: Introductionmentioning

confidence: 95%

“…Indeed, we detected the SPCA-related pump in the Golgi apparatus of E. histolytica (Rodríguez et al, 2018 ) and the SERCA-related pump in the endoplasmic reticulum of E. histolytica and E. invadens (Martinez-Higuera et al, 2013 ; Martínez-Higuera et al, 2015 ). In addition, we demonstrated that specific inhibitors of SERCA affected the encystation of E. invadens (Martínez-Higuera et al, 2015 ), suggesting that calcium flux through SERCA is involved in the development of Entamoeba sp. However, other proteins involved in the Ca 2+ movement, such as channels or exchangers, and their role in the Enatamoeba biology have not been described.…”

Section: Introductionmentioning

confidence: 95%

“…The cytolytic activity of this parasite is dependent on both, extracellular and free cytoplasmic Ca 2+ (Ravdin et al, 1982 , 1985 ). In addition, Ca 2+ flux participates in the adherence of trophozoites to fibronectin (Carbajal et al, 1996 ), as well as in growth and differentiation of E. histolytica and E. invadens (Makioka et al, 2001 , 2002 ; Martínez-Higuera et al, 2015 ). However, little is known about the proteins that regulate the Ca 2+ flux in this parasite.…”

Section: Introductionmentioning

confidence: 99%

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A Calcium/Cation Exchanger Participates in the Programmed Cell Death and in vitro Virulence of Entamoeba histolytica

Valle-Solis

Bolaños

Orozco

et al. 2018

Front. Cell. Infect. Microbiol.

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Entamoeba histolytica is the etiologic agent of human amoebiasis, disease that causes 40,000 to 100,000 deaths annually worldwide. The cytopathic activity as well as the growth and differentiation of this microorganism is dependent on both, extracellular and free cytoplasmic calcium. However, few is known about the proteins that regulate the calcium flux in this parasite. In many cells, the calcium extrusion from the cytosol is performed by plasma membrane Ca2+-ATPases and calcium/cation exchangers. The aim of this work was to identify a calcium/cation exchanger of E. histolytica and to analyze its possible role in some cellular processes triggered by calcium flux, such as the programmed cell death and in vitro virulence. By searching putative calcium/cation exchangers in the genome database of E. histolyica we identified a protein belonging to the CCX family (EhCCX). We generated a specific antibody against EhCCX, which showed that this protein was expressed in higher levels in E. histolytica than its orthologous in the non-pathogenic amoeba E. dispar. In addition, the expression of EhCCX was increased in trophozoites incubated with hydrogen peroxide. This E. histolytica exchanger was localized in the plasma membrane and in the membrane of some cytoplasmic vesicles. However, after 10 min of erythrophagocytosis, EhCCX was found predominantly in the plasma membrane of the trophozoites. On the other hand, the parasites that overexpress this exchanger contained higher cytosolic calcium levels than control, but the extrusion of calcium after the addition of hydrogen peroxide was more efficient in EhCCX-overexpressing trophozoites; consequently, the programmed cell death was retarded in these parasites. Interestingly, the overexpression of EhCCX increased the in vitro virulence of trophozoites. These results suggest that EhCCX plays important roles in the programmed cell death and in the in vitro virulence of E. histolytica.

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Entamoeba invadens: Identification of a SERCA protein and effect of SERCA inhibitors on encystation

Cited by 12 publications

References 35 publications

Targeting Channels and Transporters in Protozoan Parasite Infections

Targeting Channels and Transporters in Protozoan Parasite Infections

The NAD+ Responsive Transcription Factor ERM-BP Functions Downstream of Cellular Aggregation and Is an Early Regulator of Development and Heat Shock Response in Entamoeba

A Calcium/Cation Exchanger Participates in the Programmed Cell Death and in vitro Virulence of Entamoeba histolytica

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