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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Autor principal: Schoijet, Alejandra Cecilia
Publicado: 2017
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Acceso en línea: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
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spelling 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
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