Intracellular survival of apicomplexan parasites and host cell modification

ABSTRACT Ca2+-dependent exocytosis is essential for the life cycle of apicomplexan parasites. Toxoplasma gondii harbors a phosphoglucomutase (PGM) ortholog, PRP1, previously associated with Ca2+-dependent microneme secretion. Here it is shown that genetic deletion of either PRP1, its PGM2 ortholog, or both genes is dispensable for the parasite’s lytic cycle, including host cell egress and invasion. Depletion of the proteins abrogated high Ca2+-mediated microneme secretion induced by the ionophore A23187; however, the constitutive and phosphatidic acid-mediated release remained unaffected. Secretion mediated by the former pathway is not essential for tachyzoite survival or acute in vivo infection in the mice. Paralogs of the widely prevalent phosphoglucomutase (PGM) protein called parafusin function in calcium (Ca2+)-mediated exocytosis across eukaryotes. In Toxoplasma gondii, the parafusin-related protein 1 (PRP1) has been associated with Ca2+-dependent microneme organelle secretion required for essential processes like host cell invasion and egress. Using reverse genetics, we observed PRP1 to be dispensable for completion of the lytic cycle, including host cell invasion and egress by the parasite. However, the absence of the gene affected increased microneme release triggered by A23187, a Ca2+ ionophore used to raise the cytoplasmic Ca2+ concentration mimicking the physiological role of Ca2+ during invasion and egress. The basal levels of constitutive microneme release in extracellular parasites and phosphatidic acid-triggered microneme secretion were unaffected in the mutant. The phenotype of the deletion mutant of the second PGM-encoding gene in Toxoplasma, PGM2, was similar to the phenotype of the PRP1 deletion mutant. Furthermore, the ability of the tachyzoites to induce acute infection in the mice remained normal in the absence of both PGM paralogs. Our data thus reveal that the microneme secretion upon high Ca2+ flux is facilitated by the Toxoplasma PGM paralogs, PRP1 and PGM2. However, this protein-mediated release is neither essential for lytic cycle completion nor for acute virulence of the parasite. IMPORTANCE Ca2+-dependent exocytosis is essential for the life cycle of apicomplexan parasites. Toxoplasma gondii harbors a phosphoglucomutase (PGM) ortholog, PRP1, previously associated with Ca2+-dependent microneme secretion. Here it is shown that genetic deletion of either PRP1, its PGM2 ortholog, or both genes is dispensable for the parasite’s lytic cycle, including host cell egress and invasion. Depletion of the proteins abrogated high Ca2+-mediated microneme secretion induced by the ionophore A23187; however, the constitutive and phosphatidic acid-mediated release remained unaffected. Secretion mediated by the former pathway is not essential for tachyzoite survival or acute in vivo infection in the mice.

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Intracellular Gene Expression Profile of Listeria monocytogenes

Infection and Immunity ◽

10.1128/iai.74.2.1323-1338.2006 ◽

2006 ◽

Vol 74 (2) ◽

pp. 1323-1338 ◽

Cited By ~ 258

Author(s):

Som Subhra Chatterjee ◽

Hamid Hossain ◽

Sonja Otten ◽

Carsten Kuenne ◽

Katja Kuchmina ◽

...

Keyword(s):

Gene Expression ◽

Listeria Monocytogenes ◽

Host Cell ◽

Gene Expression Profile ◽

Expression Profile ◽

Intracellular Survival ◽

Host Cells ◽

Wall Structure ◽

Glucose Limitation ◽

Serotype 4B

ABSTRACT Listeria monocytogenes is a gram-positive, food-borne microorganism responsible for invasive infections with a high overall mortality. L. monocytogenes is among the very few microorganisms that can induce uptake into the host cell and subsequently enter the host cell cytosol by breaching the vacuolar membrane. We infected the murine macrophage cell line P388D1 with L. monocytogenes strain EGD-e and examined the gene expression profile of L. monocytogenes inside the vacuolar and cytosolic environments of the host cell by using whole-genome microarray and mutant analyses. We found that ∼17% of the total genome was mobilized to enable adaptation for intracellular growth. Intracellularly expressed genes showed responses typical of glucose limitation within bacteria, with a decrease in the amount of mRNA encoding enzymes in the central metabolism and a temporal induction of genes involved in alternative-carbon-source utilization pathways and their regulation. Adaptive intracellular gene expression involved genes that are associated with virulence, the general stress response, cell division, and changes in cell wall structure and included many genes with unknown functions. A total of 41 genes were species specific, being absent from the genome of the nonpathogenic Listeria innocua CLIP 11262 strain. We also detected 25 genes that were strain specific, i.e., absent from the genome of the previously sequenced L. monocytogenes F2365 serotype 4b strain, suggesting heterogeneity in the gene pool required for intracellular survival of L. monocytogenes in host cells. Overall, our study provides crucial insights into the strategy of intracellular survival and measures taken by L. monocytogenes to escape the host cell responses.

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Apicomplexans pulling the strings: manipulation of the host cell cytoskeleton dynamics

Parasitology ◽

10.1017/s0031182016000524 ◽

2016 ◽

Vol 143 (8) ◽

pp. 957-970 ◽

Cited By ~ 6

Author(s):

RITA CARDOSO ◽

HELENA SOARES ◽

ANDREW HEMPHILL ◽

ALEXANDRE LEITÃO

Keyword(s):

Host Cell ◽

Life Cycle Stage ◽

Effector Proteins ◽

Cell Physiology ◽

Apicomplexan Parasites ◽

Cell Cytoskeleton ◽

Cytoskeletal Filament ◽

Associated Proteins ◽

Organelle Trafficking ◽

Cytoskeleton Dynamics

SUMMARYInvasive stages of apicomplexan parasites require a host cell to survive, proliferate and advance to the next life cycle stage. Once invasion is achieved, apicomplexans interact closely with the host cell cytoskeleton, but in many cases the different species have evolved distinct mechanisms and pathways to modulate the structural organization of cytoskeletal filaments. The host cell cytoskeleton is a complex network, largely, but not exclusively, composed of microtubules, actin microfilaments and intermediate filaments, all of which are modulated by associated proteins, and it is involved in diverse functions including maintenance of cell morphology and mechanical support, migration, signal transduction, nutrient uptake, membrane and organelle trafficking and cell division. The ability of apicomplexans to modulate the cytoskeleton to their own advantage is clearly beneficial. We here review different aspects of the interactions of apicomplexans with the three main cytoskeletal filament types, provide information on the currently known parasite effector proteins and respective host cell targets involved, and how these interactions modulate the host cell physiology. Some of these findings could provide novel targets that could be exploited for the development of preventive and/or therapeutic strategies.

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Apicomplexan parasites and subversion of the host cell microRNA pathway

Trends in Parasitology ◽

10.1016/j.pt.2011.07.001 ◽

2011 ◽

Vol 27 (11) ◽

pp. 481-486 ◽

Cited By ~ 34

Author(s):

Mohamed-ali Hakimi ◽

Dominique Cannella

Keyword(s):

Host Cell ◽

Apicomplexan Parasites ◽

Microrna Pathway

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Type IV Secretion Machinery Promotes Ton-Independent Intracellular Survival of Neisseria gonorrhoeae within Cervical Epithelial Cells

Infection and Immunity ◽

10.1128/iai.00228-10 ◽

2010 ◽

Vol 78 (6) ◽

pp. 2429-2437 ◽

Cited By ~ 14

Author(s):

Tracey A. Zola ◽

Heather R. Strange ◽

Nadia M. Dominguez ◽

Joseph P. Dillard ◽

Cynthia N. Cornelissen

Keyword(s):

Epithelial Cells ◽

Neisseria Gonorrhoeae ◽

Host Cell ◽

Iron Acquisition ◽

Intracellular Survival ◽

Type Iv Secretion ◽

Open Reading Frames ◽

Cell Infection ◽

Type Iv ◽

Cervical Epithelial Cells

ABSTRACT Survival of Neisseria gonorrhoeae within host epithelial cells is expected to be important in the pathogenesis of gonococcal disease. We previously demonstrated that strain FA1090 derives iron from a host cell in a process that requires the Ton complex and a putative TonB-dependent transporter, TdfF. FA1090, however, lacks the gonococcal genetic island (GGI) that is present in the majority of strains. The GGI in strain MS11 has been partially characterized, and it encodes a type IV secretion system (T4SS) involved in DNA release. In this study we investigated the role of iron acquisition and GGI-encoded gene products in gonococcal survival within cervical epithelial cells. We demonstrated that intracellular survival of MS11 was dependent on acquisition of iron from the host cell, but unlike the findings for FA1090, expression of the Ton complex was not required. Survival was not dependent on a putative TonB-like protein encoded in the GGI but instead was directly linked to T4SS structural components in a manner independent of the ability to release or internalize DNA. These data suggest that expression of selected GGI-encoded open reading frames confers an advantage during cervical cell infection. This study provides the first link between expression of the T4SS apparatus and intracellular survival of gonococci.

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A Toxoplasma gondii Ortholog of Plasmodium GAMA Contributes to Parasite Attachment and Cell Invasion

mSphere ◽

10.1128/msphere.00012-16 ◽

2016 ◽

Vol 1 (1) ◽

Cited By ~ 10

Author(s):

My-Hang Huynh ◽

Vern B. Carruthers

Keyword(s):

Toxoplasma Gondii ◽

Host Cell ◽

Cell Invasion ◽

Genetic Evidence ◽

Specific Role ◽

Host Cells ◽

Human Pathogen ◽

Content Type ◽

Apicomplexan Parasites ◽

Adhesive Proteins

ABSTRACT Toxoplasma gondii is a successful human pathogen in the same phylum as malaria-causing Plasmodium parasites. Invasion of a host cell is an essential process that begins with secretion of adhesive proteins onto the parasite surface for attachment and subsequent penetration of the host cell. Conserved invasion proteins likely play roles that were maintained through the divergence of these parasites. Here, we identify a new conserved invasion protein called glycosylphosphatidylinositol-anchored micronemal antigen (GAMA). Tachyzoites lacking TgGAMA were partially impaired in parasite attachment and invasion of host cells, yielding the first genetic evidence of a specific role in parasite entry into host cells. These findings widen our appreciation of the repertoire of conserved proteins that apicomplexan parasites employ for cell invasion. Toxoplasma gondii and its Plasmodium kin share a well-conserved invasion process, including sequential secretion of adhesive molecules for host cell attachment and invasion. However, only a few orthologs have been shown to be important for efficient invasion by both genera. Bioinformatic screening to uncover potential new players in invasion identified a previously unrecognized T. gondii ortholog of Plasmodium glycosylphosphatidylinositol-anchored micronemal antigen (TgGAMA). We show that TgGAMA localizes to the micronemes and is processed into several proteolytic products within the parasite prior to secretion onto the parasite surface during invasion. TgGAMA from parasite lysate bound to several different host cell types in vitro, suggesting a role in parasite attachment. Consistent with this function, tetracycline-regulatable TgGAMA and TgGAMA knockout strains showed significant reductions in host cell invasion at the attachment step, with no defects in any of the other stages of the parasite lytic cycle. Together, the results of this work reveal a new conserved component of the adhesive repertoire of apicomplexan parasites. IMPORTANCE Toxoplasma gondii is a successful human pathogen in the same phylum as malaria-causing Plasmodium parasites. Invasion of a host cell is an essential process that begins with secretion of adhesive proteins onto the parasite surface for attachment and subsequent penetration of the host cell. Conserved invasion proteins likely play roles that were maintained through the divergence of these parasites. Here, we identify a new conserved invasion protein called glycosylphosphatidylinositol-anchored micronemal antigen (GAMA). Tachyzoites lacking TgGAMA were partially impaired in parasite attachment and invasion of host cells, yielding the first genetic evidence of a specific role in parasite entry into host cells. These findings widen our appreciation of the repertoire of conserved proteins that apicomplexan parasites employ for cell invasion.

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Reassessing the mechanics of parasite motility and host-cell invasion

The Journal of Cell Biology ◽

10.1083/jcb.201605100 ◽

2016 ◽

Vol 214 (5) ◽

pp. 507-515 ◽

Cited By ~ 42

Author(s):

Isabelle Tardieux ◽

Jake Baum

Keyword(s):

Plasma Membrane ◽

Host Cell ◽

Cell Invasion ◽

Motor System ◽

Current Model ◽

Cell Movement ◽

Host Cell Invasion ◽

Motor Functions ◽

Apicomplexan Parasites ◽

Parasite Motility

The capacity to migrate is fundamental to multicellular and single-celled life. Apicomplexan parasites, an ancient protozoan clade that includes malaria parasites (Plasmodium) and Toxoplasma, achieve remarkable speeds of directional cell movement. This rapidity is achieved via a divergent actomyosin motor system, housed within a narrow compartment that lies underneath the length of the parasite plasma membrane. How this motor functions at a mechanistic level during motility and host cell invasion is a matter of debate. Here, we integrate old and new insights toward refining the current model for the function of this motor with the aim of revitalizing interest in the mechanics of how these deadly pathogens move.

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Not a Simple Tether: Binding of Toxoplasma gondii AMA1 to RON2 during Invasion Protects AMA1 from Rhomboid-Mediated Cleavage and Leads to Dephosphorylation of Its Cytosolic Tail

mBio ◽

10.1128/mbio.00754-16 ◽

2016 ◽

Vol 7 (5) ◽

Cited By ~ 13

Author(s):

Shruthi Krishnamurthy ◽

Bin Deng ◽

Roxana del Rio ◽

Kerry R. Buchholz ◽

Moritz Treeck ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Host Cell ◽

Cell Invasion ◽

Transmembrane Domain ◽

Critical Role ◽

Host Cells ◽

Receptor Protein ◽

Host Cell Invasion ◽

Cell Binding ◽

Apicomplexan Parasites

ABSTRACT Apical membrane antigen 1 (AMA1) is a receptor protein on the surface of Toxoplasma gondii that plays a critical role in host cell invasion. The ligand to which T . gondii AMA1 (TgAMA1) binds, TgRON2, is secreted into the host cell membrane by the parasite during the early stages of invasion. The TgAMA1-TgRON2 complex forms the core of the “moving junction,” a ring-shaped zone of tight contact between the parasite and host cell membranes, through which the parasite pushes itself during invasion. Paradoxically, the parasite also expresses rhomboid proteases that constitutively cleave the TgAMA1 transmembrane domain. How can TgAMA1 function effectively in host cell binding if its extracellular domain is constantly shed from the parasite surface? We show here that when TgAMA1 binds the domain 3 (D3) peptide of TgRON2, its susceptibility to cleavage by rhomboid protease(s) is greatly reduced. This likely serves to maintain parasite-host cell binding at the moving junction, a hypothesis supported by data showing that parasites expressing a hypercleavable version of TgAMA1 invade less efficiently than wild-type parasites do. Treatment of parasites with the D3 peptide was also found to reduce phosphorylation of S527 on the cytoplasmic tail of TgAMA1, and parasites expressing a phosphomimetic S527D allele of TgAMA1 showed an invasion defect. Taken together, these data suggest that TgAMA1-TgRON2 interaction at the moving junction protects TgAMA1 molecules that are actively engaged in host cell penetration from rhomboid-mediated cleavage and generates an outside-in signal that leads to dephosphorylation of the TgAMA1 cytosolic tail. Both of these effects are required for maximally efficient host cell invasion. IMPORTANCE Nearly one-third of the world’s population is infected with the protozoan parasite Toxoplasma gondii , which causes life-threatening disease in neonates and immunocompromised individuals. T. gondii is a member of the phylum Apicomplexa, which includes many other parasites of veterinary and medical importance, such as those that cause coccidiosis, babesiosis, and malaria. Apicomplexan parasites grow within their hosts through repeated cycles of host cell invasion, parasite replication, and host cell lysis. Parasites that cannot invade host cells cannot survive or cause disease. AMA1 is a highly conserved protein on the surface of apicomplexan parasites that is known to be important for invasion, and the work presented here reveals new and unexpected insights into AMA1 function. A more complete understanding of the role of AMA1 in invasion may ultimately contribute to the development of new chemotherapeutics designed to disrupt AMA1 function and invasion-related signaling in this important group of human pathogens.

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Monocyte-derived chicken macrophages exposed to Eimeria tenella sporozoites display reduced susceptibility to invasion by Toxoplasma gondii tachyzoites

10.21203/rs.3.rs-21822/v1 ◽

2020 ◽

Author(s):

Runhui Zhang ◽

Wanpeng Zheng ◽

Arwid Daugschies ◽

Berit Bangoura

Keyword(s):

Host Cell ◽

Cell Invasion ◽

Cell Attachment ◽

Gliding Motility ◽

Phagocytic Cells ◽

Apicomplexan Parasites ◽

Cell Responses ◽

Host Cell Attachment ◽

Parasite Replication ◽

Host Parasite

Abstract Background: Both Eimeria (E.) tenella and Toxoplasma (T.) gondii are common apicomplexan parasites in chickens. Host cell invasion by both protozoans includes gliding motility, host cell attachment and active penetration. Chicken macrophages as phagocytic cells participate in the innate host immune response against these two parasites. Methods: In this study, primary chicken monocyte-derived macrophages (MMs) were infected with both pathogens to investigate mutual and host-parasite interactions. MMs cultures were assigned to groups that were infected with E. tenella, T. gondii, or both. In co-infected cultures, MMs were first exposed to E. tenella sporozoites for 2 hours. Afterwards, T. gondii tachyzoite infection was performed. Live-cell imaging was carried out to observe cell invasion and survival of T. gondii by single parasite tracking over a period of 20 hours post infection (hpi). Quantitative analysis for parasite replication was performed at 2, 6, 12 and 24 hpi by real-time PCR (qPCR).Results: We found that during early co-infection T. gondii tachyzoites adhered for more than 4 hours to macrophages. Although they displayed high motility, ability to penetrate the cell membrane of the potential host cell was reduced. qPCR results confirmed that significantly less T. gondii entered E. tenella-activated MMs at 2 hpi, and a reduced proportion of intracellular T. gondii survived and replicated in these cells at 24 hpi. Conclusions: We conclude that E. tenella modulates host cell responses to another apicomplexan agent, T. gondii, reducing active invasion and multiplication in chicken primary macrophages.

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