Cargando…

5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub

Hypersensitive site 5 (5′HS5) of the β-globin Locus Control Region functions as a developmental stage-specific border in erythroid cells. Here, we have analyzed the role of 5′HS5 in the three dimensional organization of the β-gene locus using the Chromatin Conformation Capture (3C) technique. The re...

Descripción completa

Detalles Bibliográficos
Autores principales: Chan, Ping Kei, Wai, Albert, Philipsen, Sjaak, Tan-Un, Kian-Cheng
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2358975/
https://www.ncbi.nlm.nih.gov/pubmed/18461170
http://dx.doi.org/10.1371/journal.pone.0002134
_version_ 1782152876604260352
author Chan, Ping Kei
Wai, Albert
Philipsen, Sjaak
Tan-Un, Kian-Cheng
author_facet Chan, Ping Kei
Wai, Albert
Philipsen, Sjaak
Tan-Un, Kian-Cheng
author_sort Chan, Ping Kei
collection PubMed
description Hypersensitive site 5 (5′HS5) of the β-globin Locus Control Region functions as a developmental stage-specific border in erythroid cells. Here, we have analyzed the role of 5′HS5 in the three dimensional organization of the β-gene locus using the Chromatin Conformation Capture (3C) technique. The results show that when 5′HS5 is deleted from the locus, both remote and internal regulatory elements are still able to interact with each other in a three-dimensional configuration termed the Active Chromatin Hub. Thus, the absence of 5′HS5 does not have an appreciable effect on the three dimensional organization of the β-globin locus. This rules out models in which 5′HS5 nucleates interactions with remote and/or internal regulatory elements. We also determined the binding of CTCF, the only defined insulator protein in mammalian cells, to 5′HS5 by using chromatin immunoprecipitation (ChIP) assays. We detect low levels of CTCF binding to 5′HS5 in primitive erythroid cells, in which it functions as a border element. Surprisingly, we also observe binding levels of CTCF to 5′HS5 in definitive erythroid cells. Thus, binding of CTCF to 5′HS5 per se does not render it a functional border element. This is consistent with the previous data suggesting that CTCF has dual functionality.
format Text
id pubmed-2358975
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-23589752008-05-07 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub Chan, Ping Kei Wai, Albert Philipsen, Sjaak Tan-Un, Kian-Cheng PLoS One Research Article Hypersensitive site 5 (5′HS5) of the β-globin Locus Control Region functions as a developmental stage-specific border in erythroid cells. Here, we have analyzed the role of 5′HS5 in the three dimensional organization of the β-gene locus using the Chromatin Conformation Capture (3C) technique. The results show that when 5′HS5 is deleted from the locus, both remote and internal regulatory elements are still able to interact with each other in a three-dimensional configuration termed the Active Chromatin Hub. Thus, the absence of 5′HS5 does not have an appreciable effect on the three dimensional organization of the β-globin locus. This rules out models in which 5′HS5 nucleates interactions with remote and/or internal regulatory elements. We also determined the binding of CTCF, the only defined insulator protein in mammalian cells, to 5′HS5 by using chromatin immunoprecipitation (ChIP) assays. We detect low levels of CTCF binding to 5′HS5 in primitive erythroid cells, in which it functions as a border element. Surprisingly, we also observe binding levels of CTCF to 5′HS5 in definitive erythroid cells. Thus, binding of CTCF to 5′HS5 per se does not render it a functional border element. This is consistent with the previous data suggesting that CTCF has dual functionality. Public Library of Science 2008-05-07 /pmc/articles/PMC2358975/ /pubmed/18461170 http://dx.doi.org/10.1371/journal.pone.0002134 Text en Chan 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chan, Ping Kei
Wai, Albert
Philipsen, Sjaak
Tan-Un, Kian-Cheng
5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title_full 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title_fullStr 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title_full_unstemmed 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title_short 5′HS5 of the Human β-globin Locus Control Region Is Dispensable for the Formation of the β-globin Active Chromatin Hub
title_sort 5′hs5 of the human β-globin locus control region is dispensable for the formation of the β-globin active chromatin hub
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2358975/
https://www.ncbi.nlm.nih.gov/pubmed/18461170
http://dx.doi.org/10.1371/journal.pone.0002134
work_keys_str_mv AT chanpingkei 5hs5ofthehumanbglobinlocuscontrolregionisdispensablefortheformationofthebglobinactivechromatinhub
AT waialbert 5hs5ofthehumanbglobinlocuscontrolregionisdispensablefortheformationofthebglobinactivechromatinhub
AT philipsensjaak 5hs5ofthehumanbglobinlocuscontrolregionisdispensablefortheformationofthebglobinactivechromatinhub
AT tanunkiancheng 5hs5ofthehumanbglobinlocuscontrolregionisdispensablefortheformationofthebglobinactivechromatinhub