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Abstract:

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

Registro:

Documento: Artículo
Título:The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi
Autor:Schoijet, A.C.; Sternlieb, T.; Alonso, G.D.
Filiación:Instituto de Investigaciones en Ingeniería Genética y Biología Molecular “Dr. Héctor N. Torres”, Vuelta de Obligado 2490, Buenos Aires, C1428ADN, Argentina
Departamento de Fisiología, Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Buenos Aires, C1428EGA CABA, Argentina
Palabras clave: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
Año:2017
Volumen:64
Número:3
Página de inicio:308
Página de fin:321
DOI: http://dx.doi.org/10.1111/jeu.12367
Título revista:Journal of Eukaryotic Microbiology
Título revista abreviado:J. Eukaryotic Microbiol.
ISSN:10665234
CODEN:JEMIE
CAS:cadaverine, 1476-39-7, 462-94-2; dansylcadaverine, 10121-91-2; phosphatidylinositol 3 kinase, 115926-52-8; Cadaverine; Class III Phosphatidylinositol 3-Kinases; DNA, Protozoan; monodansylcadaverine; Phosphatidylinositol 3-Kinases; Protozoan Proteins; Vacuolar Sorting Protein VPS15
Registro:http://digital.bl.fcen.uba.ar/collection/paper/document/paper_10665234_v64_n3_p308_Schoijet

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Citas:

---------- APA ----------
Schoijet, A.C., Sternlieb, T. & Alonso, G.D. (2017) . The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi. Journal of Eukaryotic Microbiology, 64(3), 308-321.
http://dx.doi.org/10.1111/jeu.12367
---------- CHICAGO ----------
Schoijet, A.C., Sternlieb, T., Alonso, G.D. "The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi" . Journal of Eukaryotic Microbiology 64, no. 3 (2017) : 308-321.
http://dx.doi.org/10.1111/jeu.12367
---------- MLA ----------
Schoijet, A.C., Sternlieb, T., Alonso, G.D. "The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi" . Journal of Eukaryotic Microbiology, vol. 64, no. 3, 2017, pp. 308-321.
http://dx.doi.org/10.1111/jeu.12367
---------- VANCOUVER ----------
Schoijet, A.C., Sternlieb, T., Alonso, G.D. The Phosphatidylinositol 3-kinase Class III Complex Containing TcVps15 and TcVps34 Participates in Autophagy in Trypanosoma cruzi. J. Eukaryotic Microbiol. 2017;64(3):308-321.
http://dx.doi.org/10.1111/jeu.12367