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Transcriptional Auto-Regulation of RUNX1 P1 Promoter

RUNX1 a member of the family of runt related transcription factors (RUNX), is essential for hematopoiesis. The expression of RUNX1 gene is controlled by two promoters; the distal P1 promoter and the proximal P2 promoter. Several isoforms of RUNX1 mRNA are generated through the use of both promoters...

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Autores principales: Martinez, Milka, Hinojosa, Marcela, Trombly, Daniel, Morin, Violeta, Stein, Janet, Stein, Gary, Javed, Amjad, Gutierrez, Soraya E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762634/
https://www.ncbi.nlm.nih.gov/pubmed/26901859
http://dx.doi.org/10.1371/journal.pone.0149119
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author Martinez, Milka
Hinojosa, Marcela
Trombly, Daniel
Morin, Violeta
Stein, Janet
Stein, Gary
Javed, Amjad
Gutierrez, Soraya E.
author_facet Martinez, Milka
Hinojosa, Marcela
Trombly, Daniel
Morin, Violeta
Stein, Janet
Stein, Gary
Javed, Amjad
Gutierrez, Soraya E.
author_sort Martinez, Milka
collection PubMed
description RUNX1 a member of the family of runt related transcription factors (RUNX), is essential for hematopoiesis. The expression of RUNX1 gene is controlled by two promoters; the distal P1 promoter and the proximal P2 promoter. Several isoforms of RUNX1 mRNA are generated through the use of both promoters and alternative splicing. These isoforms not only differs in their temporal expression pattern but also exhibit differences in tissue specificity. The RUNX1 isoforms derived from P2 are expressed in a variety of tissues, but expression of P1-derived isoform is restricted to cells of hematopoietic lineage. However, the control of hematopoietic-cell specific expression is poorly understood. Here we report regulation of P1-derived RUNX1 mRNA by RUNX1 protein. In silico analysis of P1 promoter revealed presence of two evolutionary conserved RUNX motifs, 0.6kb upstream of the transcription start site, and three RUNX motifs within 170bp of the 5’UTR. Transcriptional contribution of these RUNX motifs was studied in myeloid and T-cells. RUNX1 genomic fragment containing all sites show very low basal activity in both cell types. Mutation or deletion of RUNX motifs in the UTR enhances basal activity of the RUNX1 promoter. Chromatin immunoprecipitation revealed that RUNX1 protein is recruited to these sites. Overexpression of RUNX1 in non-hematopoietic cells results in a dose dependent activation of the RUNX1 P1 promoter. We also demonstrate that RUNX1 protein regulates transcription of endogenous RUNX1 mRNA in T-cell. Finally we show that SCL transcription factor is recruited to regions containing RUNX motifs in the promoter and the UTR and regulates activity of the RUNX1 P1 promoter in vitro. Thus, multiple lines of evidence show that RUNX1 protein regulates its own gene transcription.
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spelling pubmed-47626342016-03-07 Transcriptional Auto-Regulation of RUNX1 P1 Promoter Martinez, Milka Hinojosa, Marcela Trombly, Daniel Morin, Violeta Stein, Janet Stein, Gary Javed, Amjad Gutierrez, Soraya E. PLoS One Research Article RUNX1 a member of the family of runt related transcription factors (RUNX), is essential for hematopoiesis. The expression of RUNX1 gene is controlled by two promoters; the distal P1 promoter and the proximal P2 promoter. Several isoforms of RUNX1 mRNA are generated through the use of both promoters and alternative splicing. These isoforms not only differs in their temporal expression pattern but also exhibit differences in tissue specificity. The RUNX1 isoforms derived from P2 are expressed in a variety of tissues, but expression of P1-derived isoform is restricted to cells of hematopoietic lineage. However, the control of hematopoietic-cell specific expression is poorly understood. Here we report regulation of P1-derived RUNX1 mRNA by RUNX1 protein. In silico analysis of P1 promoter revealed presence of two evolutionary conserved RUNX motifs, 0.6kb upstream of the transcription start site, and three RUNX motifs within 170bp of the 5’UTR. Transcriptional contribution of these RUNX motifs was studied in myeloid and T-cells. RUNX1 genomic fragment containing all sites show very low basal activity in both cell types. Mutation or deletion of RUNX motifs in the UTR enhances basal activity of the RUNX1 promoter. Chromatin immunoprecipitation revealed that RUNX1 protein is recruited to these sites. Overexpression of RUNX1 in non-hematopoietic cells results in a dose dependent activation of the RUNX1 P1 promoter. We also demonstrate that RUNX1 protein regulates transcription of endogenous RUNX1 mRNA in T-cell. Finally we show that SCL transcription factor is recruited to regions containing RUNX motifs in the promoter and the UTR and regulates activity of the RUNX1 P1 promoter in vitro. Thus, multiple lines of evidence show that RUNX1 protein regulates its own gene transcription. Public Library of Science 2016-02-22 /pmc/articles/PMC4762634/ /pubmed/26901859 http://dx.doi.org/10.1371/journal.pone.0149119 Text en © 2016 Martinez et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Martinez, Milka
Hinojosa, Marcela
Trombly, Daniel
Morin, Violeta
Stein, Janet
Stein, Gary
Javed, Amjad
Gutierrez, Soraya E.
Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title_full Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title_fullStr Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title_full_unstemmed Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title_short Transcriptional Auto-Regulation of RUNX1 P1 Promoter
title_sort transcriptional auto-regulation of runx1 p1 promoter
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762634/
https://www.ncbi.nlm.nih.gov/pubmed/26901859
http://dx.doi.org/10.1371/journal.pone.0149119
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