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PICOT binding to the polycomb group protein, EED, alters H3K27 methylation at the MYT1 PRC2 target gene.
Pandya, Pinakin; Pasvolsky, Ronit; Babichev, Yael; Braiman, Alex; Witte, Stephan; Altman, Amnon; Isakov, Noah.
Afiliación
  • Pandya P; The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences and the Cancer Research Center, Ben Gurion University of the Negev, P.O.B. 653, Beer Sheva, 84105, Israel.
  • Pasvolsky R; The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences and the Cancer Research Center, Ben Gurion University of the Negev, P.O.B. 653, Beer Sheva, 84105, Israel.
  • Babichev Y; The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences and the Cancer Research Center, Ben Gurion University of the Negev, P.O.B. 653, Beer Sheva, 84105, Israel.
  • Braiman A; The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences and the Cancer Research Center, Ben Gurion University of the Negev, P.O.B. 653, Beer Sheva, 84105, Israel.
  • Witte S; Division of Cell Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
  • Altman A; Division of Cell Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
  • Isakov N; The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences and the Cancer Research Center, Ben Gurion University of the Negev, P.O.B. 653, Beer Sheva, 84105, Israel. Electronic address: noah@bgu.ac.il.
Biochem Biophys Res Commun ; 509(2): 469-475, 2019 02 05.
Article en En | MEDLINE | ID: mdl-30595380
PICOT is a ubiquitous protein that has no functional redundant ortholog and is critical for mouse embryonic development. It is involved in the regulation of signal transduction in T lymphocytes and cardiac muscle, and in cellular iron metabolism and biogenesis of Fe/S proteins. However, very little is known about the physiological role of PICOT and its mechanism of action, and on its upstream regulators or downstream target molecules. In attempt to identify new PICOT interaction partners, we adopted the yeast two-hybrid system and screened a Jurkat T cell cDNA library using the full-length human PICOT cDNA as a bait. We found that PICOT interacts with embryonic ectoderm development (EED), a Polycomb Group (PcG) protein that serves as a core component of the Polycomb repressive complex 2 (PRC2) and contributes to the regulation of chromatin remodeling and cell differentiation. Using bead immobilized GST-PICOT and GST-EED fusion proteins in a pull-down assay and reciprocal coimmunoprecipitation studies we demonstrated that the interaction between PICOT and EED also occurs in human Jurkat T cells. In addition, immunofluorescence staining of Jurkat T cells revealed partial colocalization of PICOT and EED, predominantly in the cell nuclei. A pull-down assay using the GST-EED fusion protein and lysates of cells expressing different Myc-tagged truncation products of PICOT revealed that binding of EED is mediated by each of the two C-terminal PICOT homology domains and suggests that simultaneous interaction via both domains increases the binding affinity. Furthermore, PICOT knock-down in Jurkat T cells resulted in a reduced histone H3 lysine 27 trimethylation (H3K27me3) at the PRC2 target gene, myelin transcription factor 1 (MYT1), suggesting that PICOT binding to EED alters PRC2-regulated transcriptional repression, and potentially contributes to the epigenetic regulation of chromatin silencing and remodeling.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factores de Transcripción / Histonas / Proteínas Portadoras / Regiones Promotoras Genéticas / Proteínas de Unión al ADN / Complejo Represivo Polycomb 2 Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2019 Tipo del documento: Article País de afiliación: Israel Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factores de Transcripción / Histonas / Proteínas Portadoras / Regiones Promotoras Genéticas / Proteínas de Unión al ADN / Complejo Represivo Polycomb 2 Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2019 Tipo del documento: Article País de afiliación: Israel Pais de publicación: Estados Unidos