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Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-270527

RESUMO

<p><b>OBJECTIVE</b>The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase, the so-called TgPP2C which has been suggested to contribute to parasite growth regulation. Ectopic expression in mammalian cells also indicated that the enzyme could regulate growth and survival. In this study, we aimed to investigate the interaction of TgPP2C with human SSRP1 (structure-specific recognition protein 1) and the effects of TgPP2C on cell viability.</p><p><b>METHODS</b>The yeast two hybrid system, His-tag pull-down and co-immunoprecipitation assays were used to confirm the interaction of TgPP2C with SSRP1 and determine the binding domain on SSRP1. The evaluation of cell apoptosis was performed using cleaved caspase-3 antibody and Annexin-V/PI kit combined with flow cytometry.</p><p><b>RESULTS</b>We identified human SSRP1 as an interacting partner of TgPP2C. The C-terminal region of SSRP1 including the amino acids 471 to 538 was specifically mapped as the region responsible for interaction with TgPP2C. The overexpression of TgPP2C down-regulated cell apoptosis and negatively regulated apoptosis induced by DRB, casein kinase II (CKII) inhibitor, through enhanced interaction with SSRP1.</p><p><b>CONCLUSION</b>TgPP2C may be a parasitic factor capable of promoting cell survival through interaction with the host protein SSRP1, thereby creating a favorable environment for parasite growth.</p>


Assuntos
Humanos , Apoptose , Western Blotting , Proteínas de Ligação a DNA , Genética , Metabolismo , Citometria de Fluxo , Células HeLa , Proteínas de Grupo de Alta Mobilidade , Genética , Metabolismo , Imunoprecipitação , Fosfoproteínas Fosfatases , Genética , Metabolismo , Proteína Fosfatase 2C , Toxoplasma , Fatores de Elongação da Transcrição , Genética , Metabolismo , Técnicas do Sistema de Duplo-Híbrido
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