5'HS5 of the human beta-globin locus control region is dispensable for the formation of the beta-globin active chromatin hub

Sep 16, 2026

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Hypersensitive site 5 (5'HS5) of the β-globin Locus Control Region serves as a developmental stage-specific border in erythroid cells. This study uses Chromatin Conformation Capture (3C) to test whether 5'HS5 is required for the three-dimensional organization of the β-gene locus. Deletion of 5'HS5 preserves interactions among remote and internal regulatory elements, maintaining an Active Chromatin Hub configuration. Consequently, 5'HS5 does not appreciably influence locus 3D structure. Binding of CTCF to 5'HS5 is measured by ChIP, showing low binding in primitive erythroid cells and unexpected binding in definitive erythroid cells, indicating that CTCF binding alone is insufficient for border function.

LanguageEnglish

CategoryGeneral

FormatPDF

Pages10

Reading Time4 min

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Abstract

Hypersensitive site 5 (5'HS5) and border function

3C analysis of three-dimensional locus organization

CTCF binding assessed by ChIP assays

Introduction

EUR Research Information Portal 5'HS5 of the human beta-globin locus control region is dispensable for the formation of the beta-globin active chromatin hub Published in: PLoS One (print) Publication status and date: Published: 01/01/2008 DOI (link to publisher): 10.1371/journal.pone.0002134 Document Version Publisher's PDF, also known as Version of record Citation for the published version (APA): Chan, PK. , Wai, A. , Philipsen, S. , & Tan-Un, KC. (2008) . 5'HS5 of the human beta-globin locus control region is dispensable for the formation of the beta-globin active chromatin hub. PLoS One (print) , 3 , e2134 . [https://doi.org/10.1371/journal.pone.0002134](https://doi.org/10.1371/journal.pone.0002134) Link to publication on the EUR Research Information Portal Terms and Conditions of Use Except as permitted by the applicable copyright law, you may not reproduce or make this material available to any third party without the prior written permission from the copyright holder(s) . Copyright law allows the following uses of this material without prior permission: • you may download, save and print a copy of this material for your personal use only; • you may share the EUR portal link to this material. In case the material is published with an open access license (e.g. a Creative Commons (CC) license), other uses may be allowed. Please check the terms and conditions of the specific license. Take-down policy If you believe that this material infringes your copyright and/or any other intellectual property rights, you may request its removal by contacting us at the following email address: [openaccess.library@eur.nl. Please](openaccess.library@eur.nl. Please) provide us with all the relevant information, including the reasons why you believe any of your rights have been infringed. In case of a legitimate complaint, we will make the material inaccessible and/or remove it from the website. 5 9 HS5 of the Human b-globin Locus Control Region Is Dispensable for the Formation of the b-globin Active Chromatin Hub Ping Kei Chan 1, Albert Wai2, Sjaak Philipsen2*, Kian-Cheng Tan-Un 1* 1 Department of Zoology, Kadoorie Biological Science Building, The University of Hong Kong, Hong Kong Special Administrative Region (SAR), China, 2Erasmus MC, Department of Cell Biology, Rotterdam, The Netherlands | Abstract
Hypersensitive site 5 (5 9HS5) of the b-globin Locus Control Region functions as a developmental stage-specific border inerythroid cells. Here, we have analyzed the role of 5 9HS5 in the three dimensional organization of the b-gene locus using the Chromatin Conformation Capture (3C) technique. The results show that when 5 9HS5 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 9HS5 does not have an appreciable effect on the three dimensional organization of the b-globin locus. This rules out models in which 5 9HS5 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 9HS5 by using chromatin immunoprecipitation (ChIP) assays. We detect low levels of CTCF binding to 5 9HS5 in primitive erythroid cells, in which it functions as a border element. Surprisingly, we also observe binding levels of CTCF to 5 9HS5 in definitive erythroid cells. Thus, binding of CTCF to 5 9HS5 per se does not render it a functional border element. This is consistent with the previous data suggesting that CTCF has dual functionality. | | --- | | Citation: Chan PK, Wai A, Philipsen S, Tan-Un K-C (2008) 5 9HS5 of the Human b-globin Locus Control Region Is Dispensable for the Formation of the b-globin
Active Chromatin Hub. PLoS ONE 3(5): e2134 . doi:10.1371/journal.pone.0002134
Editor: Axel Imhof, University of Munich and Center of Integrated Prote

FAQs

What role does 5'HS5 play in the β-globin locus under normal conditions?

5'HS5 functions as a developmental stage-specific border element in erythroid cells within the β-globin Locus Control Region.

How does deleting 5'HS5 affect the three-dimensional organization of the β-globin locus?

Deletion of 5'HS5 does not measurably disrupt the 3D organization; remote and internal regulatory elements still interact in an Active Chromatin Hub configuration.

Does CTCF binding to 5'HS5 automatically make it a functional border element?

No. The study finds CTCF binding to 5'HS5 in both primitive and definitive erythroid cells, yet binding alone does not guarantee border functionality.

Sep 16, 2026

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