scholarly journals Fibronectin-Degrading Activity of Trypanosoma cruzi Cysteine Proteinase Plays a Role in Host Cell Invasion

2014 ◽  
Vol 82 (12) ◽  
pp. 5166-5174 ◽  
Author(s):  
Fernando Yukio Maeda ◽  
Cristian Cortez ◽  
Mario Augusto Izidoro ◽  
Luiz Juliano ◽  
Nobuko Yoshida
Keyword(s):  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Hela Cells ◽  
Transforming Growth Factor ◽  
Cysteine Proteinase ◽  
Mammalian Host ◽  
Cysteine Proteinase Inhibitor ◽  
Host Cell Invasion ◽  
Content Type

ABSTRACTTrypanosoma cruzi, the agent of Chagas disease, binds to diverse extracellular matrix proteins. Such an ability prevails in the parasite forms that circulate in the bloodstream and contributes to host cell invasion. Whether this also applies to the insect-stage metacyclic trypomastigotes, the developmental forms that initiate infection in the mammalian host, is not clear. UsingT. cruziCL strain metacyclic forms, we investigated whether fibronectin bound to the parasites and affected target cell invasion. Fibronectin present in cell culture medium bound to metacyclic forms and was digested by cruzipain, the majorT. cruzicysteine proteinase. G strain, with negligible cruzipain activity, displayed a minimal fibronectin-degrading effect. Binding to fibronectin was mediated by gp82, the metacyclic stage-specific surface molecule implicated in parasite internalization. When exogenous fibronectin was present at concentrations higher than cruzipain can properly digest, or fibronectin expression was stimulated by treatment of epithelial HeLa cells with transforming growth factor beta, the parasite invasion was reduced. Treatment of HeLa cells with purified recombinant cruzipain increased parasite internalization, whereas the treatment of parasites with cysteine proteinase inhibitor had the opposite effect. Metacyclic trypomastigote entry into HeLa cells was not affected by anti-β1 integrin antibody but was inhibited by anti-fibronectin antibody. Overall, our results have indicated that the cysteine proteinase ofT. cruzimetacyclic forms, through its fibronectin-degrading activity, is implicated in host cell invasion.

scholarly journals Parasite-Mediated Remodeling of the Host Microfilament Cytoskeleton Enables Rapid Egress of Trypanosoma cruzi following Membrane Rupture

mBio ◽  
2021 ◽  
Author(s):  
Eden R. Ferreira ◽  
Alexis Bonfim-Melo ◽  
Barbara A. Burleigh ◽  
Jaime A. Costales ◽  
Kevin M. Tyler ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Lytic Cycle ◽  
Host Cells ◽  
Host Cell Invasion ◽  
High Quality ◽  
Content Type ◽  
Membrane Rupture

Understanding how Trypanosoma cruzi interacts with host cells has been transformed by high-quality studies that have examined in detail the mechanisms of T. cruzi host cell invasion. In contrast, little is known about the latter stages of the parasite’s lytic cycle: how parasites egress and thereby sustain round after round of infection.

scholarly journals Molecular Characterization of a Novel Family of Trypanosoma cruzi Surface Membrane Proteins (TcSMP) Involved in Mammalian Host Cell Invasion

2015 ◽  
Vol 9 (11) ◽  
pp. e0004216 ◽  
Author(s):  
Nadini Oliveira Martins ◽  
Renata Torres de Souza ◽  
Esteban Mauricio Cordero ◽  
Danielle Cortez Maldonado ◽  
Cristian Cortez ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Molecular Characterization ◽  
Cell Invasion ◽  
Surface Membrane ◽  
Mammalian Host ◽  
Host Cell Invasion

scholarly journals Actin Cytoskeleton-Dependent and -Independent Host Cell Invasion by Trypanosoma cruzi Is Mediated by Distinct Parasite Surface Molecules

2006 ◽  
Vol 74 (10) ◽  
pp. 5522-5528 ◽  
Author(s):  
Daniele Ferreira ◽  
Mauro Cortez ◽  
Vanessa D. Atayde ◽  
Nobuko Yoshida
Keyword(s):  
Trypanosoma Cruzi ◽  
Actin Cytoskeleton ◽  
Host Cell ◽  
Cell Invasion ◽  
Hela Cells ◽  
Cytochalasin D ◽  
Target Cells ◽  
Host Cell Invasion ◽  
Latrunculin B ◽  
Surface Molecules

ABSTRACT The disassembly of host cell actin cytoskeleton as a facilitator of Trypanosoma cruzi invasion has been reported by some authors, while other workers claim that it instead inhibits internalization of the parasite. In this study we aimed at elucidating the basis of this discrepancy. We performed experiments with metacyclic trypomastigotes of T. cruzi strains G and CL, which differ markedly in infectivity and enter target cells by engaging the surface molecules gp35/50 and gp82, respectively, which have signaling activity. Treatment of HeLa cells with the F-actin-disrupting drug cytochalasin D or latrunculin B inhibited the invasion by strain G but not the invasion by strain CL. In contrast to cells penetrated by strain CL, which were previously shown to have a disrupted actin cytoskeleton architecture, no such alteration was observed in HeLa cells invaded by strain G, and parasites were found to be closely associated with target cell actin. Coinfection with enteroinvasive Escherichia coli (EIEC), which recruits host cell actin for internalization, drastically reduced entry of strain CL into HeLa cells but not entry of strain G. In contrast to gp82 in its recombinant form, which induces disruption of F-actin and inhibits EIEC invasion, purified mucin-like gp35/50 molecules promoted an increase in EIEC uptake by HeLa cells. These data, plus the finding that drugs that interfere with mammalian cell signaling differentially affect the internalization of metacyclic forms of strains G and CL, indicate that the host cell invasion mediated by gp35/50 is associated with signaling events that favor actin recruitment, in contrast to gp82-dependent invasion, which triggers the signaling pathways leading to disassembly of F-actin.

scholarly journals Deletion of a Golgi protein in Trypanosoma cruzi reveals a critical role for Mn2+ in protein glycosylation needed for host cell invasion and intracellular replication

2021 ◽  
Vol 17 (3) ◽  
pp. e1009399
Author(s):  
Srinivasan Ramakrishnan ◽  
Linn M. Unger ◽  
Rodrigo P. Baptista ◽  
Teresa Cruz-Bustos ◽  
Roberto Docampo
Keyword(s):  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Critical Role ◽  
Protein Glycosylation ◽  
Growth Defect ◽  
Mammalian Host ◽  
Host Cell Invasion ◽  
Intracellular Replication ◽  
Parasite Life Cycle

Trypanosoma cruzi is a protist parasite and the causative agent of American trypanosomiasis or Chagas disease. The parasite life cycle in its mammalian host includes an intracellular stage, and glycosylated proteins play a key role in host-parasite interaction facilitating adhesion, invasion and immune evasion. Here, we report that a Golgi-localized Mn2+-Ca2+/H+ exchanger of T. cruzi (TcGDT1) is required for efficient protein glycosylation, host cell invasion, and intracellular replication. The Golgi localization was determined by immunofluorescence and electron microscopy assays. TcGDT1 was able to complement the growth defect of Saccharomyces cerevisiae null mutants of its ortholog ScGDT1 but ablation of TcGDT1 by CRISPR/Cas9 did not affect the growth of the insect stage of the parasite. The defect in protein glycosylation was rescued by Mn2+ supplementation to the growth medium, underscoring the importance of this transition metal for Golgi glycosylation of proteins.

scholarly journals Protein Kinase A Is Essential for Invasion of Plasmodium falciparum into Human Erythrocytes

mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Mary-Louise Wilde ◽  
Tony Triglia ◽  
Danushka Marapana ◽  
Jennifer K. Thompson ◽  
Alexei A. Kouzmitchev ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Host Cell ◽  
Cell Invasion ◽  
Blood Cells ◽  
Host Cell Invasion ◽  
Parasite Invasion ◽  
Content Type ◽  
Kinase A

ABSTRACT Understanding the mechanisms behind host cell invasion by Plasmodium falciparum remains a major hurdle to developing antimalarial therapeutics that target the asexual cycle and the symptomatic stage of malaria. Host cell entry is enabled by a multitude of precisely timed and tightly regulated receptor-ligand interactions. Cyclic nucleotide signaling has been implicated in regulating parasite invasion, and an important downstream effector of the cAMP-signaling pathway is protein kinase A (PKA), a cAMP-dependent protein kinase. There is increasing evidence that P. falciparum PKA (PfPKA) is responsible for phosphorylation of the cytoplasmic domain of P. falciparum apical membrane antigen 1 (PfAMA1) at Ser610, a cAMP-dependent event that is crucial for successful parasite invasion. In the present study, CRISPR-Cas9 and conditional gene deletion (dimerizable cre) technologies were implemented to generate a P. falciparum parasite line in which expression of the catalytic subunit of PfPKA (PfPKAc) is under conditional control, demonstrating highly efficient dimerizable Cre recombinase (DiCre)-mediated gene excision and complete knockdown of protein expression. Parasites lacking PfPKAc show severely reduced growth after one intraerythrocytic growth cycle and are deficient in host cell invasion, as highlighted by live-imaging experiments. Furthermore, PfPKAc-deficient parasites are unable to phosphorylate PfAMA1 at Ser610. This work not only identifies an essential role for PfPKAc in the P. falciparum asexual life cycle but also confirms that PfPKAc is the kinase responsible for phosphorylating PfAMA1 Ser610. IMPORTANCE Malaria continues to present a major global health burden, particularly in low-resource countries. Plasmodium falciparum, the parasite responsible for the most severe form of malaria, causes disease through rapid and repeated rounds of invasion and replication within red blood cells. Invasion into red blood cells is essential for P. falciparum survival, and the molecular events mediating this process have gained much attention as potential therapeutic targets. With no effective vaccine available, and with the emergence of resistance to antimalarials, there is an urgent need for the development of new therapeutics. Our research has used genetic techniques to provide evidence of an essential protein kinase involved in P. falciparum invasion. Our work adds to the current understanding of parasite signaling processes required for invasion, highlighting PKA as a potential drug target to inhibit invasion for the treatment of malaria.

scholarly journals Inhibition of Host Cell Lysosome Spreading by Trypanosoma cruzi Metacyclic Stage-Specific Surface Molecule gp90 Downregulates Parasite Invasion

2017 ◽  
Vol 85 (9) ◽  
Author(s):  
João Paulo Ferreira Rodrigues ◽  
Guilherme Hideki Takahashi Sant'ana ◽  
Maria Aparecida Juliano ◽  
Nobuko Yoshida
Keyword(s):  
Trypanosoma Cruzi ◽  
Hela Cell ◽  
Specific Surface ◽  
Host Cell ◽  
Cell Invasion ◽  
Target Cell ◽  
Content Type ◽  
Terminal Domain ◽  
Successful Infection

ABSTRACT Successful infection by Trypanosoma cruzi, the agent of Chagas' disease, is critically dependent on host cell invasion by metacyclic trypomastigote (MT) forms. Two main metacyclic stage-specific surface molecules, gp82 and gp90, play determinant roles in target cell invasion in vitro and in oral T. cruzi infection in mice. The structure and properties of gp82, which is highly conserved among T. cruzi strains, are well known. Information on gp90 is still rather sparse. Here, we attempted to fill that gap. gp90, purified from poorly invasive G strain MT and expressing gp90 at high levels, inhibited HeLa cell lysosome spreading and the gp82-mediated internalization of a highly invasive CL strain MT expressing low levels of a diverse gp90 molecule. A recombinant protein containing the conserved C-terminal domain of gp90 exhibited the same properties as the native G strain gp90: it counteracted the host cell lysosome spreading induced by recombinant gp82 and exhibited an inhibitory effect on HeLa cell invasion by CL strain MT. Assays to identify the gp90 sequence associated with the property of downregulating MT invasion, using synthetic peptides spanning the gp90 C-terminal domain, revealed the sequence GVLYTADKEW. These data, plus the findings that lysosome spreading was induced upon HeLa cell interaction with CL strain MT, but not with G strain MT, and that in mixed infection CL strain MT internalization was inhibited by G strain MT, suggest that the inhibition of target cell lysosome spreading is the mechanism by which the gp90 molecule exerts its downregulatory role.

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