The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi
Autophagy is a degradative process by which eukaryotic cells digest their own components to provide aminoacids that may function as energy source under nutritional stress conditions. There is experimental evidence for autophagy in parasitic protists belonging to the family Trypanosomatidae. However,...
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paper:paper_10665234_v64_n3_p308_Schoijet2023-06-08T16:04:19Z The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi Schoijet, Alejandra Cecilia Autophagy Trypanosoma cruzi vesicular acidification Vps15 Vps34 amino acid biochemical composition catalysis cells and cell components chemotherapy enzyme enzyme activity gene expression membrane parasite Eukaryota Trypanosoma cruzi Trypanosomatidae cadaverine dansylcadaverine phosphatidylinositol 3 kinase protein serine threonine kinase VPS15 protozoal DNA protozoal protein analogs and derivatives animal autophagy biosynthesis cell culture technique cell membrane cell vacuole cytology enzyme assay enzymology gene expression regulation genetic transfection genetics life cycle stage metabolism molecular cloning phagosome physiology sequence analysis site directed mutagenesis Trypanosoma cruzi two hybrid system Animals Autophagy Cadaverine Cell Culture Techniques Cell Membrane Class III Phosphatidylinositol 3-Kinases Cloning, Molecular DNA, Protozoan Enzyme Assays Gene Expression Regulation, Enzymologic Life Cycle Stages Mutagenesis, Site-Directed Phagosomes Phosphatidylinositol 3-Kinases Protozoan Proteins Sequence Analysis Transfection Trypanosoma cruzi Two-Hybrid System Techniques Vacuolar Sorting Protein VPS15 Vacuoles Autophagy is a degradative process by which eukaryotic cells digest their own components to provide aminoacids that may function as energy source under nutritional stress conditions. There is experimental evidence for autophagy in parasitic protists belonging to the family Trypanosomatidae. However, few proteins implicated in this process have been characterized so far in these parasites. Moreover, it has been shown that autophagy is involved in Trypanosoma cruzi differentiation and thus might have a role in pathogenicity. Here, we report the cloning and biochemical characterization of TcVps15. In addition, we demonstrate that TcVps15 interact with the PI3K TcVps34 and that both proteins associate with cellular membranes. Under nutritional stress conditions, TcVps15 and TcVps34 modify their subcellular distribution showing a partial co-localization in autophagosomes with TcAtg8.1 and using an active site TcVps15-mutated version (TcVps15-K219D-HA) we demonstrated that this relocalization depends on the TcVps15 catalytic activity. Overexpression of TcVps15-HA and TcVps15-K219D-HA also leads to increased accumulation of monodansylcadaverine (MDC) in autophagic vacuoles under nutritional stress conditions compared to wild-type cells. In addition, the MDC-specific activity shows to be significantly higher in TcVps15-HA overexpressing cells when compared with TcVps15-K219D-HA. Our results reveal for the first time a role of TcVps15 as a key regulator of TcVps34 enzymatic activity and implicate the TcVps15-Vps34 complex in autophagy in T. cruzi, exposing a new key pathway to explore novel chemotherapeutic targets. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists Fil:Schoijet, A.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2017 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10665234_v64_n3_p308_Schoijet http://hdl.handle.net/20.500.12110/paper_10665234_v64_n3_p308_Schoijet |
institution |
Universidad de Buenos Aires |
institution_str |
I-28 |
repository_str |
R-134 |
collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
topic |
Autophagy Trypanosoma cruzi vesicular acidification Vps15 Vps34 amino acid biochemical composition catalysis cells and cell components chemotherapy enzyme enzyme activity gene expression membrane parasite Eukaryota Trypanosoma cruzi Trypanosomatidae cadaverine dansylcadaverine phosphatidylinositol 3 kinase protein serine threonine kinase VPS15 protozoal DNA protozoal protein analogs and derivatives animal autophagy biosynthesis cell culture technique cell membrane cell vacuole cytology enzyme assay enzymology gene expression regulation genetic transfection genetics life cycle stage metabolism molecular cloning phagosome physiology sequence analysis site directed mutagenesis Trypanosoma cruzi two hybrid system Animals Autophagy Cadaverine Cell Culture Techniques Cell Membrane Class III Phosphatidylinositol 3-Kinases Cloning, Molecular DNA, Protozoan Enzyme Assays Gene Expression Regulation, Enzymologic Life Cycle Stages Mutagenesis, Site-Directed Phagosomes Phosphatidylinositol 3-Kinases Protozoan Proteins Sequence Analysis Transfection Trypanosoma cruzi Two-Hybrid System Techniques Vacuolar Sorting Protein VPS15 Vacuoles |
spellingShingle |
Autophagy Trypanosoma cruzi vesicular acidification Vps15 Vps34 amino acid biochemical composition catalysis cells and cell components chemotherapy enzyme enzyme activity gene expression membrane parasite Eukaryota Trypanosoma cruzi Trypanosomatidae cadaverine dansylcadaverine phosphatidylinositol 3 kinase protein serine threonine kinase VPS15 protozoal DNA protozoal protein analogs and derivatives animal autophagy biosynthesis cell culture technique cell membrane cell vacuole cytology enzyme assay enzymology gene expression regulation genetic transfection genetics life cycle stage metabolism molecular cloning phagosome physiology sequence analysis site directed mutagenesis Trypanosoma cruzi two hybrid system Animals Autophagy Cadaverine Cell Culture Techniques Cell Membrane Class III Phosphatidylinositol 3-Kinases Cloning, Molecular DNA, Protozoan Enzyme Assays Gene Expression Regulation, Enzymologic Life Cycle Stages Mutagenesis, Site-Directed Phagosomes Phosphatidylinositol 3-Kinases Protozoan Proteins Sequence Analysis Transfection Trypanosoma cruzi Two-Hybrid System Techniques Vacuolar Sorting Protein VPS15 Vacuoles Schoijet, Alejandra Cecilia The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
topic_facet |
Autophagy Trypanosoma cruzi vesicular acidification Vps15 Vps34 amino acid biochemical composition catalysis cells and cell components chemotherapy enzyme enzyme activity gene expression membrane parasite Eukaryota Trypanosoma cruzi Trypanosomatidae cadaverine dansylcadaverine phosphatidylinositol 3 kinase protein serine threonine kinase VPS15 protozoal DNA protozoal protein analogs and derivatives animal autophagy biosynthesis cell culture technique cell membrane cell vacuole cytology enzyme assay enzymology gene expression regulation genetic transfection genetics life cycle stage metabolism molecular cloning phagosome physiology sequence analysis site directed mutagenesis Trypanosoma cruzi two hybrid system Animals Autophagy Cadaverine Cell Culture Techniques Cell Membrane Class III Phosphatidylinositol 3-Kinases Cloning, Molecular DNA, Protozoan Enzyme Assays Gene Expression Regulation, Enzymologic Life Cycle Stages Mutagenesis, Site-Directed Phagosomes Phosphatidylinositol 3-Kinases Protozoan Proteins Sequence Analysis Transfection Trypanosoma cruzi Two-Hybrid System Techniques Vacuolar Sorting Protein VPS15 Vacuoles |
description |
Autophagy is a degradative process by which eukaryotic cells digest their own components to provide aminoacids that may function as energy source under nutritional stress conditions. There is experimental evidence for autophagy in parasitic protists belonging to the family Trypanosomatidae. However, few proteins implicated in this process have been characterized so far in these parasites. Moreover, it has been shown that autophagy is involved in Trypanosoma cruzi differentiation and thus might have a role in pathogenicity. Here, we report the cloning and biochemical characterization of TcVps15. In addition, we demonstrate that TcVps15 interact with the PI3K TcVps34 and that both proteins associate with cellular membranes. Under nutritional stress conditions, TcVps15 and TcVps34 modify their subcellular distribution showing a partial co-localization in autophagosomes with TcAtg8.1 and using an active site TcVps15-mutated version (TcVps15-K219D-HA) we demonstrated that this relocalization depends on the TcVps15 catalytic activity. Overexpression of TcVps15-HA and TcVps15-K219D-HA also leads to increased accumulation of monodansylcadaverine (MDC) in autophagic vacuoles under nutritional stress conditions compared to wild-type cells. In addition, the MDC-specific activity shows to be significantly higher in TcVps15-HA overexpressing cells when compared with TcVps15-K219D-HA. Our results reveal for the first time a role of TcVps15 as a key regulator of TcVps34 enzymatic activity and implicate the TcVps15-Vps34 complex in autophagy in T. cruzi, exposing a new key pathway to explore novel chemotherapeutic targets. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists |
author |
Schoijet, Alejandra Cecilia |
author_facet |
Schoijet, Alejandra Cecilia |
author_sort |
Schoijet, Alejandra Cecilia |
title |
The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
title_short |
The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
title_full |
The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
title_fullStr |
The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
title_full_unstemmed |
The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi |
title_sort |
phosphatidylinositol 3-kinase class iii complex containing tcvps15 and tcvps34 participates in autophagy in trypanosoma cruzi |
publishDate |
2017 |
url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10665234_v64_n3_p308_Schoijet http://hdl.handle.net/20.500.12110/paper_10665234_v64_n3_p308_Schoijet |
work_keys_str_mv |
AT schoijetalejandracecilia thephosphatidylinositol3kinaseclassiiicomplexcontainingtcvps15andtcvps34participatesinautophagyintrypanosomacruzi AT schoijetalejandracecilia phosphatidylinositol3kinaseclassiiicomplexcontainingtcvps15andtcvps34participatesinautophagyintrypanosomacruzi |
_version_ |
1768546128213573632 |