Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.

Somatic activating mutations in the Notch1 receptor result in the overexpression of activated Notch1, which can be tumorigenic. The goal of this study is to understand the molecular mechanisms underlying the phenotypic changes caused by the overexpression of ligand independent Notch 1 by using a tet...

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Autores principales:	Habib Hamidi, Derek Gustafason, Matteo Pellegrini, Judith Gasson
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e9af31b0b88347be9fbf64ddf4e7014a
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spelling	oai:doaj.org-article:e9af31b0b88347be9fbf64ddf4e7014a2021-11-18T06:53:09ZIdentification of novel targets of CSL-dependent Notch signaling in hematopoiesis.1932-620310.1371/journal.pone.0020022https://doaj.org/article/e9af31b0b88347be9fbf64ddf4e7014a2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21637838/?tool=EBIhttps://doaj.org/toc/1932-6203Somatic activating mutations in the Notch1 receptor result in the overexpression of activated Notch1, which can be tumorigenic. The goal of this study is to understand the molecular mechanisms underlying the phenotypic changes caused by the overexpression of ligand independent Notch 1 by using a tetracycline inducible promoter in an in vitro embryonic stem (ES) cells/OP9 stromal cells coculture system, recapitulating normal hematopoiesis. First, an in silico analysis of the promoters of Notch regulated genes (previously determined by microarray analysis) revealed that the motifs recognized by regulatory proteins known to mediate hematopoiesis were overrepresented. Notch 1 does not bind DNA but instead binds the CSL transcription factor to regulate gene expression. The in silico analysis also showed that there were putative CSL binding sites observed in the promoters of 28 out of 148 genes. A custom ChIP-chip array was used to assess the occupancy of CSL in the promoter regions of the Notch1 regulated genes in vivo and showed that 61 genes were bound by activated Notch responsive CSL. Then, comprehensive mapping of the CSL binding sites genome-wide using ChIP-seq analysis revealed that over 10,000 genes were bound within 10 kb of the TSS (transcription start site). The majority of the targets discovered by ChIP-seq belong to pathways that have been shown by others to crosstalk with Notch signaling. Finally, 83 miRNAs were significantly differentially expressed by greater than 1.5-fold during the course of in vitro hematopoiesis. Thirty one miRNA were up-regulated and fifty two were down-regulated. Overexpression of Notch1 altered this pattern of expression of microRNA: six miRNAs were up-regulated and four were down regulated as a result of activated Notch1 overexpression during the course of hematopoiesis. Time course analysis of hematopoietic development revealed that cells with Notch 1 overexpression mimic miRNA expression of cells in a less mature stage, which is consistent with our previous biological characterization.Habib HamidiDerek GustafasonMatteo PellegriniJudith GassonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 5, p e20022 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Habib Hamidi Derek Gustafason Matteo Pellegrini Judith Gasson Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
description	Somatic activating mutations in the Notch1 receptor result in the overexpression of activated Notch1, which can be tumorigenic. The goal of this study is to understand the molecular mechanisms underlying the phenotypic changes caused by the overexpression of ligand independent Notch 1 by using a tetracycline inducible promoter in an in vitro embryonic stem (ES) cells/OP9 stromal cells coculture system, recapitulating normal hematopoiesis. First, an in silico analysis of the promoters of Notch regulated genes (previously determined by microarray analysis) revealed that the motifs recognized by regulatory proteins known to mediate hematopoiesis were overrepresented. Notch 1 does not bind DNA but instead binds the CSL transcription factor to regulate gene expression. The in silico analysis also showed that there were putative CSL binding sites observed in the promoters of 28 out of 148 genes. A custom ChIP-chip array was used to assess the occupancy of CSL in the promoter regions of the Notch1 regulated genes in vivo and showed that 61 genes were bound by activated Notch responsive CSL. Then, comprehensive mapping of the CSL binding sites genome-wide using ChIP-seq analysis revealed that over 10,000 genes were bound within 10 kb of the TSS (transcription start site). The majority of the targets discovered by ChIP-seq belong to pathways that have been shown by others to crosstalk with Notch signaling. Finally, 83 miRNAs were significantly differentially expressed by greater than 1.5-fold during the course of in vitro hematopoiesis. Thirty one miRNA were up-regulated and fifty two were down-regulated. Overexpression of Notch1 altered this pattern of expression of microRNA: six miRNAs were up-regulated and four were down regulated as a result of activated Notch1 overexpression during the course of hematopoiesis. Time course analysis of hematopoietic development revealed that cells with Notch 1 overexpression mimic miRNA expression of cells in a less mature stage, which is consistent with our previous biological characterization.
format	article
author	Habib Hamidi Derek Gustafason Matteo Pellegrini Judith Gasson
author_facet	Habib Hamidi Derek Gustafason Matteo Pellegrini Judith Gasson
author_sort	Habib Hamidi
title	Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
title_short	Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
title_full	Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
title_fullStr	Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
title_full_unstemmed	Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.
title_sort	identification of novel targets of csl-dependent notch signaling in hematopoiesis.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/e9af31b0b88347be9fbf64ddf4e7014a
work_keys_str_mv	AT habibhamidi identificationofnoveltargetsofcsldependentnotchsignalinginhematopoiesis AT derekgustafason identificationofnoveltargetsofcsldependentnotchsignalinginhematopoiesis AT matteopellegrini identificationofnoveltargetsofcsldependentnotchsignalinginhematopoiesis AT judithgasson identificationofnoveltargetsofcsldependentnotchsignalinginhematopoiesis
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Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis.

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