NbHSWP11, a MicrosporidiaNosema bombycisProtein, Localizing in the Spore Wall and Membranes, Reduces Spore Adherence to Host Cell BME

2014 ◽  
Vol 100 (5) ◽  
pp. 623-632 ◽  
Author(s):  
Donglin Yang ◽  
Xiaoqun Dang ◽  
Pai Peng ◽  
Mengxian Long ◽  
Cheng Ma ◽  
...  
Keyword(s):  
Host Cell ◽  
Spore Wall ◽  
Spore Adherence

scholarly journals EnP1, a Microsporidian Spore Wall Protein That Enables Spores To Adhere to and Infect Host Cells In Vitro

2007 ◽  
Vol 6 (8) ◽  
pp. 1354-1362 ◽  
Author(s):  
Timothy R. Southern ◽  
Carrie E. Jolly ◽  
Melissa E. Lester ◽  
J. Russell Hayman
Keyword(s):  
Cell Surface ◽  
Host Cell ◽  
Spore Wall ◽  
Site Directed Mutagenesis ◽  
Host Cells ◽  
Binding Motif ◽  
Heparin Binding ◽  
Cell Infection ◽  
Host Cell Surface

ABSTRACT Microsporidia are spore-forming fungal pathogens that require the intracellular environment of host cells for propagation. We have shown that spores of the genus Encephalitozoon adhere to host cell surface glycosaminoglycans (GAGs) in vitro and that this adherence serves to modulate the infection process. In this study, a spore wall protein (EnP1; Encephalitozoon cuniculi ECU01_0820) from E. cuniculi and Encephalitozoon intestinalis is found to interact with the host cell surface. Analysis of the amino acid sequence reveals multiple heparin-binding motifs, which are known to interact with extracellular matrices. Both recombinant EnP1 protein and purified EnP1 antibody inhibit spore adherence, resulting in decreased host cell infection. Furthermore, when the N-terminal heparin-binding motif is deleted by site-directed mutagenesis, inhibition of adherence is ablated. Our transmission immunoelectron microscopy reveals that EnP1 is embedded in the microsporidial endospore and exospore and is found in high abundance in the polar sac/anchoring disk region, an area from which the everting polar tube is released. Finally, by using a host cell binding assay, EnP1 is shown to bind host cell surfaces but not to those that lack surface GAGs. Collectively, these data show that given its expression in both the endospore and the exospore, EnP1 is a microsporidian cell wall protein that may function both in a structural capacity and in modulating in vitro host cell adherence and infection.

scholarly journals Interaction and Assembly of Two Novel Proteins in the Spore Wall of the Microsporidian Species Nosema bombycis and Their Roles in Adherence to and Infection of Host Cells

2015 ◽  
Vol 83 (4) ◽  
pp. 1715-1731 ◽  
Author(s):  
Donglin Yang ◽  
Guoqing Pan ◽  
Xiaoqun Dang ◽  
Yawei Shi ◽  
Chunfeng Li ◽  
...  
Keyword(s):  
Host Cell ◽  
Spore Wall ◽  
Host Cells ◽  
Scaffolding Protein ◽  
Environmental Pressures ◽  
Content Type ◽  
Nosema Bombycis ◽  
Major Function ◽  
Microsporidian Species

Microsporidia are obligate intracellular parasites with rigid spore walls that protect against various environmental pressures. Despite an extensive description of the spore wall, little is known regarding the mechanism by which it is deposited or the role it plays in cell adhesion and infection. In this study, we report the identification and characterization of two novel spore wall proteins, SWP7 and SWP9, in the microsporidian speciesNosema bombycis. SWP7 and SWP9 are mainly localized to the exospore and endospore of mature spores and the cytoplasm of sporonts, respectively. In addition, a portion of SWP9 is targeted to the spore wall of sporoblasts earlier than SWP7 is. Both SWP7 and SWP9 are specifically colocalized to the spore wall in mature spores. Furthermore, immunoprecipitation, far-Western blotting, unreduced SDS-PAGE, and yeast two-hybrid data demonstrated that SWP7 interacted with SWP9. The chitin binding assay showed that, within the total spore protein, SWP9 and SWP7 can bind to the deproteinated chitin spore coats (DCSCs) ofN. bombycis. However, binding of the recombinant protein rSWP7-His to the DCSCs is dependent on the combination of rSWP9–glutathioneS-transferase (GST) with the DCSCs. Finally, rSWP9-GST, anti-SWP9, and anti-SWP7 antibodies decreased spore adhesion and infection of the host cell. In conclusion, SWP7 and SWP9 may have important structural capacities and play significant roles in modulating host cell adherence and infectionin vitro. A possible major function of SWP9 is as a scaffolding protein that supports other proteins (such as SWP7) that form the integrated spore wall ofN. bombycis.

scholarly journals Expression and Localization of an Hsp70 Protein in the MicrosporidianEncephalitozoon cuniculi

2010 ◽  
Vol 2010 ◽  
pp. 1-6 ◽  
Author(s):  
Carrie E. Jolly ◽  
Cory A. Leonard ◽  
J. Russell Hayman
Keyword(s):  
Host Cell ◽  
Cell Attachment ◽  
Surface Protein ◽  
Surface Proteins ◽  
Spore Surface ◽  
Dna Encoding ◽  
Hsp70 Protein ◽  
Surface Protein Expression ◽  
Host Cell Attachment ◽  
Spore Adherence

Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and matureE. cuniculispores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1) did so. In future studies, the antibodies to the ‘C1’ Hsp70 will be used to delineate spore surface protein expression.

scholarly journals Branching Network of Proteinaceous Filaments within the Parasitophorous Vacuole ofEncephalitozoon cuniculiandEncephalitozoon hellem

2011 ◽  
Vol 79 (3) ◽  
pp. 1374-1385 ◽  
Author(s):  
Kaya Ghosh ◽  
Eddie Nieves ◽  
Patrick Keeling ◽  
Jean-Francois Pombert ◽  
Philipp P. Henrich ◽  
...  
Keyword(s):  
Host Cell ◽  
Parasitophorous Vacuole ◽  
Spore Wall ◽  
Human Pathogens ◽  
Polar Tube ◽  
Molecular Approaches ◽  
Resting Spore ◽  
Intracellular Parasites ◽  
Significant Pathology ◽  
Microsporidian Spore

ABSTRACTThe microsporidia are a diverse phylum of obligate intracellular parasites that infect all major animal groups and have been recognized as emerging human pathogens for which few chemotherapeutic options currently exist. These organisms infect every tissue and organ system, causing significant pathology, especially in immune-compromised populations. The microsporidian spore employs a unique infection strategy in which its contents are delivered into a host cell via the polar tube, an organelle that lies coiled within the resting spore but erupts with a force sufficient to pierce the plasma membrane of its host cell. Using biochemical and molecular approaches, we have previously identified components of the polar tube and spore wall of the Encephalitozoonidae. In this study, we employed a shotgun proteomic strategy to identify novel structural components of these organelles inEncephalitozoon cuniculi. As a result, a new component of theE. cuniculideveloping spore wall was identified. Surprisingly, using the same approach, a heretofore undescribed filamentous network within the lumen of the parasitophorous vacuole was discovered. This network was also present in the parasitophorous vacuole ofEncephalitozoon hellem. Thus, in addition to further elucidating the molecular composition of seminal organelles and revealing novel diagnostic and therapeutic targets, proteomic analysis-driven approaches exploring the spore may also uncover unknown facets of microsporidian biology.

scholarly journals Role of Sulfated Glycans in Adherence of the Microsporidian Encephalitozoon intestinalis to Host Cells In Vitro

2005 ◽  
Vol 73 (2) ◽  
pp. 841-848 ◽  
Author(s):  
J. Russell Hayman ◽  
Timothy R. Southern ◽  
Theodore E. Nash
Keyword(s):  
Cell Surface ◽  
Host Cell ◽  
Mutant Cell ◽  
Host Cells ◽  
Target Cells ◽  
Cell Infection ◽  
Encephalitozoon Intestinalis ◽  
Successful Infection ◽  
Host Cell Surface ◽  
Spore Adherence

ABSTRACT Microsporidia are obligate intracellular opportunistic protists that infect a wide variety of animals, including humans, via environmentally resistant spores. Infection requires that spores be in close proximity to host cells so that the hollow polar tube can pierce the cell membrane and inject the spore contents into the cell cytoplasm. Like other eukaryotic microbes, microsporidia may use specific mechanisms for adherence in order to achieve target cell proximity and increase the likelihood of successful infection. Our data show that Encephalitozoon intestinalis exploits sulfated glycans such as the cell surface glycosaminoglycans (GAGs) in selection of and attachment to host cells. When exogenous sulfated glycans are used as inhibitors in spore adherence assays, E. intestinalis spore adherence is reduced by as much as 88%. However, there is no inhibition when nonsulfated glycans are used, suggesting that E. intestinalis spores utilize sulfated host cell glycans in adherence. These studies were confirmed by exposure of host cells to xylopyranoside, which limits host cell surface GAGs, and sodium chlorate, which decreases surface sulfation. Spore adherence studies with CHO mutant cell lines that are deficient in either surface GAGs or surface heparan sulfate also confirmed the necessity of sulfated glycans. Furthermore, when spore adherence is inhibited, host cell infection is reduced, indicating a direct association between spore adherence and infectivity. These data show that E. intestinalis specifically adheres to target cells by way of sulfated host cell surface GAGs and that this mechanism serves to enhance infectivity.

Bacteriophages Associated with Turkey Coecums in Bluecomb-Infected and Healthy Birds

1969 ◽  
Vol 27 ◽  
pp. 226-227
Author(s):  
A. E. Ritchie
Keyword(s):  
Host Cell ◽  
Causal Relationship ◽  
Cell Cultures ◽  
Cell Interaction ◽  
Etiologic Agent ◽  
Viral Origin ◽  
Intestinal Contents ◽  
Host Cell Interaction

The cause of bluecomb disease in turkeys is unknown. Filtration of infective intestinal contents suggests a viral origin. To date, it has not been possible to isolate the etiologic agent in various cell cultures. The purpose of this work was to characterize as many virus-like entities as were recognizable in intestines of both healthy and bluecomb-infected turkeys. By a comparison of the viral populations it was hoped that some insight might be gained into the cause of this disease. Studies of turkey hemorraghic enteritis by Gross and Moore (Avian Dis. 11: 296-307, 1967) have suggested that a bacteriophage-host cell interaction may bear some causal relationship to that disease.

The Infection of Cells by Bullet-Shaped Viruses

1969 ◽  
Vol 27 ◽  
pp. 372-373
Author(s):  
Frederick A. Murphy ◽  
Alyne K. Harrison ◽  
Sylvia G. Whitfield
Keyword(s):  
Electron Microscopy ◽  
Physical Properties ◽  
Host Cell ◽  
Thin Section ◽  
Cultured Cells ◽  
Cell Infection ◽  
Thin Section Electron Microscopy ◽  
Section Electron ◽  
Section Electron Microscopy

The bullet-shaped viruses are currently classified together on the basis of similarities in virion morphology and physical properties. Biologically and ecologically the member viruses are extremely diverse. In searching for further bases for making comparisons of these agents, the nature of host cell infection, both in vivo and in cultured cells, has been explored by thin-section electron microscopy.

Host cell signalling inhibitors against helicobacter pylori

2009 ◽  
Vol 47 (05) ◽  
Author(s):  
Z Varga ◽  
Z Greff ◽  
J Pató ◽  
P Bánhegyi ◽  
L Őrfi ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Host Cell ◽  
Cell Signalling
Sign in / Sign up

Export Citation Format

Share Document