Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall.
Giardia passes through two stages during its life cycle, the trophozoite and the cyst. Cyst formation involves the synthesis of cyst wall proteins (CWPs) and the transport of CWPs into encystation-specific vesicles (ESVs). Active vesicular trafficking is essential for encystation, but the molecular...
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ftdoajarticles:oai:doaj.org/article:6a6e089b6cde4df78ce027fa0b702a08 2023-05-15T15:14:34+02:00 Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. Araceli Castillo-Romero Gloria Leon-Avila Ching C Wang Armando Perez Rangel Minerva Camacho Nuez Carlos Garcia Tovar Jorge Tonatiuh Ayala-Sumuano Juan Pedro Luna-Arias Jose Manuel Hernandez 2010-06-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0000697 https://doaj.org/article/6a6e089b6cde4df78ce027fa0b702a08 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2879372?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000697 https://doaj.org/article/6a6e089b6cde4df78ce027fa0b702a08 PLoS Neglected Tropical Diseases, Vol 4, Iss 6, p e697 (2010) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2010 ftdoajarticles https://doi.org/10.1371/journal.pntd.0000697 2022-12-31T05:31:16Z Giardia passes through two stages during its life cycle, the trophozoite and the cyst. Cyst formation involves the synthesis of cyst wall proteins (CWPs) and the transport of CWPs into encystation-specific vesicles (ESVs). Active vesicular trafficking is essential for encystation, but the molecular machinery driving vesicular trafficking remains unknown. The Rab proteins are involved in the targeting of vesicles to several intracellular compartments through their association with cytoskeletal motor proteins.In this study, we found a relationship between Rab11 and the actin cytoskeleton in CWP1 transport. Confocal microscopy showed Rab11 was distributed throughout the entire trophozoite, while in cysts it was translocated to the periphery of the cell, where it colocalized with ESVs and microfilaments. Encystation was also accompanied by changes in rab11 mRNA expression. To evaluate the role of microfilaments in encystation, the cells were treated with latrunculin A. Scanning electron microscopy showed this treatment resulted in morphological damages to encysted parasites. The intensity of fluorescence-labeled Rab11 and CWP1 in ESVs and cyst walls was reduced, and rab11 and cwp1 mRNA levels were down-regulated. Furthermore, knocking down Rab11 with a hammerhead ribozyme resulted in an up to 80% down-regulation of rab11 mRNA. Although this knockdown did not appear lethal for trophozoites and did not affect cwp1 expression during the encystation, confocal images showed CWP1 was redistributed throughout the cytosol.Our results indicate that Rab11 participates in the early and late encystation stages by regulating CWP1 localization and the actin-mediated transport of ESVs towards the periphery. In addition, alterations in the dynamics of actin affected rab11 and cwp1 expression. Our results provide new information about the molecules involved in Giardia encystation and suggest that Rab11 and actin may be useful as novel pharmacological targets. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 4 6 e697 |
institution |
Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Araceli Castillo-Romero Gloria Leon-Avila Ching C Wang Armando Perez Rangel Minerva Camacho Nuez Carlos Garcia Tovar Jorge Tonatiuh Ayala-Sumuano Juan Pedro Luna-Arias Jose Manuel Hernandez Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Giardia passes through two stages during its life cycle, the trophozoite and the cyst. Cyst formation involves the synthesis of cyst wall proteins (CWPs) and the transport of CWPs into encystation-specific vesicles (ESVs). Active vesicular trafficking is essential for encystation, but the molecular machinery driving vesicular trafficking remains unknown. The Rab proteins are involved in the targeting of vesicles to several intracellular compartments through their association with cytoskeletal motor proteins.In this study, we found a relationship between Rab11 and the actin cytoskeleton in CWP1 transport. Confocal microscopy showed Rab11 was distributed throughout the entire trophozoite, while in cysts it was translocated to the periphery of the cell, where it colocalized with ESVs and microfilaments. Encystation was also accompanied by changes in rab11 mRNA expression. To evaluate the role of microfilaments in encystation, the cells were treated with latrunculin A. Scanning electron microscopy showed this treatment resulted in morphological damages to encysted parasites. The intensity of fluorescence-labeled Rab11 and CWP1 in ESVs and cyst walls was reduced, and rab11 and cwp1 mRNA levels were down-regulated. Furthermore, knocking down Rab11 with a hammerhead ribozyme resulted in an up to 80% down-regulation of rab11 mRNA. Although this knockdown did not appear lethal for trophozoites and did not affect cwp1 expression during the encystation, confocal images showed CWP1 was redistributed throughout the cytosol.Our results indicate that Rab11 participates in the early and late encystation stages by regulating CWP1 localization and the actin-mediated transport of ESVs towards the periphery. In addition, alterations in the dynamics of actin affected rab11 and cwp1 expression. Our results provide new information about the molecules involved in Giardia encystation and suggest that Rab11 and actin may be useful as novel pharmacological targets. |
format |
Article in Journal/Newspaper |
author |
Araceli Castillo-Romero Gloria Leon-Avila Ching C Wang Armando Perez Rangel Minerva Camacho Nuez Carlos Garcia Tovar Jorge Tonatiuh Ayala-Sumuano Juan Pedro Luna-Arias Jose Manuel Hernandez |
author_facet |
Araceli Castillo-Romero Gloria Leon-Avila Ching C Wang Armando Perez Rangel Minerva Camacho Nuez Carlos Garcia Tovar Jorge Tonatiuh Ayala-Sumuano Juan Pedro Luna-Arias Jose Manuel Hernandez |
author_sort |
Araceli Castillo-Romero |
title |
Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
title_short |
Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
title_full |
Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
title_fullStr |
Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
title_full_unstemmed |
Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
title_sort |
rab11 and actin cytoskeleton participate in giardia lamblia encystation, guiding the specific vesicles to the cyst wall. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pntd.0000697 https://doaj.org/article/6a6e089b6cde4df78ce027fa0b702a08 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 4, Iss 6, p e697 (2010) |
op_relation |
http://europepmc.org/articles/PMC2879372?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000697 https://doaj.org/article/6a6e089b6cde4df78ce027fa0b702a08 |
op_doi |
https://doi.org/10.1371/journal.pntd.0000697 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
4 |
container_issue |
6 |
container_start_page |
e697 |
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1766345010824871936 |