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Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers

Differentiation into environmentally resistant cysts is required for transmission of the ubiquitous intestinal parasite Giardia lamblia. Encystation in Giardia requires the production, processing and transport of Cyst Wall Proteins (CWPs) in developmentally induced, Golgi-like, Encystation Specific...

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Autores principales: Thomas, Elizabeth B., Sutanto, Renaldo, Johnson, Richard S., Shih, Han-Wei, Alas, Germain C. M., Krtková, Jana, MacCoss, Michael J., Paredez, Alexander R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111296/
https://www.ncbi.nlm.nih.gov/pubmed/33987184
http://dx.doi.org/10.3389/fcell.2021.662945
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author Thomas, Elizabeth B.
Sutanto, Renaldo
Johnson, Richard S.
Shih, Han-Wei
Alas, Germain C. M.
Krtková, Jana
MacCoss, Michael J.
Paredez, Alexander R.
author_facet Thomas, Elizabeth B.
Sutanto, Renaldo
Johnson, Richard S.
Shih, Han-Wei
Alas, Germain C. M.
Krtková, Jana
MacCoss, Michael J.
Paredez, Alexander R.
author_sort Thomas, Elizabeth B.
collection PubMed
description Differentiation into environmentally resistant cysts is required for transmission of the ubiquitous intestinal parasite Giardia lamblia. Encystation in Giardia requires the production, processing and transport of Cyst Wall Proteins (CWPs) in developmentally induced, Golgi-like, Encystation Specific Vesicles (ESVs). Progress through this trafficking pathway can be followed by tracking CWP localization over time. However, there is no recognized system to distinguish the advancing stages of this process which can complete at variable rates depending on how encystation is induced. Here, we propose a staging system for encysting Giardia based on the morphology of CWP1-stained ESVs. We demonstrate the molecular distinctiveness of maturing ESVs at these stages by following GlRab GTPases through encystation. Previously, we established that Giardia’s sole Rho family GTPase, GlRac, associates with ESVs and has a role in regulating their maturation and the secretion of their cargo. As a proof of principle, we delineate the relationship between GlRac and ESV stages. Through proteomic studies, we identify putative interactors of GlRac that could be used as additional ESV stage markers. This staging system provides a common descriptor of ESV maturation regardless of the source of encysting cells. Furthermore, the identified set of molecular markers for ESV stages will be a powerful tool for characterizing trafficking mutants that impair ESV maturation and morphology.
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spelling pubmed-81112962021-05-12 Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers Thomas, Elizabeth B. Sutanto, Renaldo Johnson, Richard S. Shih, Han-Wei Alas, Germain C. M. Krtková, Jana MacCoss, Michael J. Paredez, Alexander R. Front Cell Dev Biol Cell and Developmental Biology Differentiation into environmentally resistant cysts is required for transmission of the ubiquitous intestinal parasite Giardia lamblia. Encystation in Giardia requires the production, processing and transport of Cyst Wall Proteins (CWPs) in developmentally induced, Golgi-like, Encystation Specific Vesicles (ESVs). Progress through this trafficking pathway can be followed by tracking CWP localization over time. However, there is no recognized system to distinguish the advancing stages of this process which can complete at variable rates depending on how encystation is induced. Here, we propose a staging system for encysting Giardia based on the morphology of CWP1-stained ESVs. We demonstrate the molecular distinctiveness of maturing ESVs at these stages by following GlRab GTPases through encystation. Previously, we established that Giardia’s sole Rho family GTPase, GlRac, associates with ESVs and has a role in regulating their maturation and the secretion of their cargo. As a proof of principle, we delineate the relationship between GlRac and ESV stages. Through proteomic studies, we identify putative interactors of GlRac that could be used as additional ESV stage markers. This staging system provides a common descriptor of ESV maturation regardless of the source of encysting cells. Furthermore, the identified set of molecular markers for ESV stages will be a powerful tool for characterizing trafficking mutants that impair ESV maturation and morphology. Frontiers Media S.A. 2021-04-27 /pmc/articles/PMC8111296/ /pubmed/33987184 http://dx.doi.org/10.3389/fcell.2021.662945 Text en Copyright © 2021 Thomas, Sutanto, Johnson, Shih, Alas, Krtková, MacCoss and Paredez. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Thomas, Elizabeth B.
Sutanto, Renaldo
Johnson, Richard S.
Shih, Han-Wei
Alas, Germain C. M.
Krtková, Jana
MacCoss, Michael J.
Paredez, Alexander R.
Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title_full Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title_fullStr Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title_full_unstemmed Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title_short Staging Encystation Progression in Giardia lamblia Using Encystation-Specific Vesicle Morphology and Associating Molecular Markers
title_sort staging encystation progression in giardia lamblia using encystation-specific vesicle morphology and associating molecular markers
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111296/
https://www.ncbi.nlm.nih.gov/pubmed/33987184
http://dx.doi.org/10.3389/fcell.2021.662945
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