Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch

Abstract The Krüppel-like factor (KLF) family of transcription factors play critical roles in haematopoiesis. KLF1, the founding member of the family, has been implicated in the control of both erythropoiesis and megakaryopoiesis. Here we describe a novel system using an artificial dominant negative...

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Autores principales:	Laura J. Norton, Samantha Hallal, Elizabeth S. Stout, Alister P. W. Funnell, Richard C. M. Pearson, Merlin Crossley, Kate G. R. Quinlan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a37d4ae2beb44fd8a42326855e39b724
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spelling	oai:doaj.org-article:a37d4ae2beb44fd8a42326855e39b7242021-12-02T16:06:28ZDirect competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch10.1038/s41598-017-03289-52045-2322https://doaj.org/article/a37d4ae2beb44fd8a42326855e39b7242017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03289-5https://doaj.org/toc/2045-2322Abstract The Krüppel-like factor (KLF) family of transcription factors play critical roles in haematopoiesis. KLF1, the founding member of the family, has been implicated in the control of both erythropoiesis and megakaryopoiesis. Here we describe a novel system using an artificial dominant negative isoform of KLF1 to investigate the role of KLF1 in the erythroid/megakaryocytic switch in vivo. We developed murine cell lines stably overexpressing a GST-KLF1 DNA binding domain fusion protein (GST-KLF1 DBD), as well as lines expressing GST only as a control. Interestingly, overexpression of GST-KLF1 DBD led to an overall reduction in erythroid features and an increase in megakaryocytic features indicative of a reduced function of endogenous KLF1. We simultaneously compared in vivo DNA occupancy of both endogenous KLF1 and GST-KLF1 DBD by ChIP qPCR. Here we found that GST-KLF1 DBD physically displaces endogenous KLF1 at a number of loci, providing novel in vivo evidence of direct competition between DNA binding proteins. These results highlight the role of KLF1 in the erythroid/megakaryocyte switch and suggest that direct competition between transcription factors with similar consensus sequences is an important mechanism in transcriptional regulation.Laura J. NortonSamantha HallalElizabeth S. StoutAlister P. W. FunnellRichard C. M. PearsonMerlin CrossleyKate G. R. QuinlanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Laura J. Norton Samantha Hallal Elizabeth S. Stout Alister P. W. Funnell Richard C. M. Pearson Merlin Crossley Kate G. R. Quinlan Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
description	Abstract The Krüppel-like factor (KLF) family of transcription factors play critical roles in haematopoiesis. KLF1, the founding member of the family, has been implicated in the control of both erythropoiesis and megakaryopoiesis. Here we describe a novel system using an artificial dominant negative isoform of KLF1 to investigate the role of KLF1 in the erythroid/megakaryocytic switch in vivo. We developed murine cell lines stably overexpressing a GST-KLF1 DNA binding domain fusion protein (GST-KLF1 DBD), as well as lines expressing GST only as a control. Interestingly, overexpression of GST-KLF1 DBD led to an overall reduction in erythroid features and an increase in megakaryocytic features indicative of a reduced function of endogenous KLF1. We simultaneously compared in vivo DNA occupancy of both endogenous KLF1 and GST-KLF1 DBD by ChIP qPCR. Here we found that GST-KLF1 DBD physically displaces endogenous KLF1 at a number of loci, providing novel in vivo evidence of direct competition between DNA binding proteins. These results highlight the role of KLF1 in the erythroid/megakaryocyte switch and suggest that direct competition between transcription factors with similar consensus sequences is an important mechanism in transcriptional regulation.
format	article
author	Laura J. Norton Samantha Hallal Elizabeth S. Stout Alister P. W. Funnell Richard C. M. Pearson Merlin Crossley Kate G. R. Quinlan
author_facet	Laura J. Norton Samantha Hallal Elizabeth S. Stout Alister P. W. Funnell Richard C. M. Pearson Merlin Crossley Kate G. R. Quinlan
author_sort	Laura J. Norton
title	Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
title_short	Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
title_full	Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
title_fullStr	Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
title_full_unstemmed	Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch
title_sort	direct competition between dna binding factors highlights the role of krüppel-like factor 1 in the erythroid/megakaryocyte switch
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a37d4ae2beb44fd8a42326855e39b724
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Direct competition between DNA binding factors highlights the role of Krüppel-like Factor 1 in the erythroid/megakaryocyte switch

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