MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein

ABSTRACT Toxoplasma gondii causes a chronic infection that affects a significant portion of the world’s population, and this latent infection is the source of reactivation of toxoplasmosis. An attribute of the slowly growing bradyzoite stage of the parasite is the formation of a cyst within infected...

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Autores principales:	Vincent Tu, Joshua Mayoral, Rama R. Yakubu, Tadakimi Tomita, Tatsuki Sugi, Bing Han, Tere Williams, Yanfen Ma, Louis M. Weiss
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	latency monoclonal antibody screen intravacuolar network MAG1 MAG2 Toxoplasma gondii Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6176377d4b5445eca9735feae7daec95
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spelling	oai:doaj.org-article:6176377d4b5445eca9735feae7daec952021-11-15T15:27:52ZMAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein10.1128/mSphere.00100-202379-5042https://doaj.org/article/6176377d4b5445eca9735feae7daec952020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00100-20https://doaj.org/toc/2379-5042ABSTRACT Toxoplasma gondii causes a chronic infection that affects a significant portion of the world’s population, and this latent infection is the source of reactivation of toxoplasmosis. An attribute of the slowly growing bradyzoite stage of the parasite is the formation of a cyst within infected cells, allowing the parasite to escape the host’s immune response. In this study, a new bradyzoite cyst matrix antigen (MAG) was identified through a hybridoma library screen. This cyst matrix antigen, matrix antigen 2 (MAG2), contains 14 tandem repeats consisting of acidic, basic, and proline residues. Immunoblotting revealed that MAG2 migrates at a level higher than its predicted molecular weight, and computational analysis showed that the structure of MAG2 is highly disordered. Cell fractionation studies indicated that MAG2 was associated with both insoluble and soluble cyst matrix material, suggesting that it interacts with the intracyst network (ICN). Examination of the kinetics of MAG2 within the cyst matrix using fluorescence recovery after photobleaching (FRAP) demonstrated that MAG2 does not readily diffuse within the cyst matrix. Kinetic studies of MAG1 demonstrated that this protein has different diffusion kinetics in tachyzoite and bradyzoite vacuoles and that its mobility is not altered in the absence of MAG2. In addition, deletion of MAG2 does not influence growth, cystogenesis, or cyst morphology. IMPORTANCE This report expands on the list of characterized Toxoplasma gondii cyst matrix proteins. Using fluorescence recovery after photobleaching (FRAP), we have shown that matrix proteins within the cyst matrix are not mainly in a mobile state, providing further evidence of how proteins behave within the cyst matrix. Understanding the proteins expressed during the bradyzoite stage of the parasite reveals how the parasite functions during chronic infection.Vincent TuJoshua MayoralRama R. YakubuTadakimi TomitaTatsuki SugiBing HanTere WilliamsYanfen MaLouis M. WeissAmerican Society for Microbiologyarticlelatencymonoclonal antibody screenintravacuolar networkMAG1MAG2Toxoplasma gondiiMicrobiologyQR1-502ENmSphere, Vol 5, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	latency monoclonal antibody screen intravacuolar network MAG1 MAG2 Toxoplasma gondii Microbiology QR1-502
spellingShingle	latency monoclonal antibody screen intravacuolar network MAG1 MAG2 Toxoplasma gondii Microbiology QR1-502 Vincent Tu Joshua Mayoral Rama R. Yakubu Tadakimi Tomita Tatsuki Sugi Bing Han Tere Williams Yanfen Ma Louis M. Weiss MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
description	ABSTRACT Toxoplasma gondii causes a chronic infection that affects a significant portion of the world’s population, and this latent infection is the source of reactivation of toxoplasmosis. An attribute of the slowly growing bradyzoite stage of the parasite is the formation of a cyst within infected cells, allowing the parasite to escape the host’s immune response. In this study, a new bradyzoite cyst matrix antigen (MAG) was identified through a hybridoma library screen. This cyst matrix antigen, matrix antigen 2 (MAG2), contains 14 tandem repeats consisting of acidic, basic, and proline residues. Immunoblotting revealed that MAG2 migrates at a level higher than its predicted molecular weight, and computational analysis showed that the structure of MAG2 is highly disordered. Cell fractionation studies indicated that MAG2 was associated with both insoluble and soluble cyst matrix material, suggesting that it interacts with the intracyst network (ICN). Examination of the kinetics of MAG2 within the cyst matrix using fluorescence recovery after photobleaching (FRAP) demonstrated that MAG2 does not readily diffuse within the cyst matrix. Kinetic studies of MAG1 demonstrated that this protein has different diffusion kinetics in tachyzoite and bradyzoite vacuoles and that its mobility is not altered in the absence of MAG2. In addition, deletion of MAG2 does not influence growth, cystogenesis, or cyst morphology. IMPORTANCE This report expands on the list of characterized Toxoplasma gondii cyst matrix proteins. Using fluorescence recovery after photobleaching (FRAP), we have shown that matrix proteins within the cyst matrix are not mainly in a mobile state, providing further evidence of how proteins behave within the cyst matrix. Understanding the proteins expressed during the bradyzoite stage of the parasite reveals how the parasite functions during chronic infection.
format	article
author	Vincent Tu Joshua Mayoral Rama R. Yakubu Tadakimi Tomita Tatsuki Sugi Bing Han Tere Williams Yanfen Ma Louis M. Weiss
author_facet	Vincent Tu Joshua Mayoral Rama R. Yakubu Tadakimi Tomita Tatsuki Sugi Bing Han Tere Williams Yanfen Ma Louis M. Weiss
author_sort	Vincent Tu
title	MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
title_short	MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
title_full	MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
title_fullStr	MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
title_full_unstemmed	MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein
title_sort	mag2, a <named-content content-type="genus-species">toxoplasma gondii</named-content> bradyzoite stage-specific cyst matrix protein
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/6176377d4b5445eca9735feae7daec95
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MAG2, a <named-content content-type="genus-species">Toxoplasma gondii</named-content> Bradyzoite Stage-Specific Cyst Matrix Protein

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