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Nucleic Acids Research Advance Access published online on July 11, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn429
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Genomics

Spatial configuration of the chicken {alpha}-globin gene domain: immature and active chromatin hubs

Alexey A. Gavrilov and Sergey V. Razin*

Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology of the Russian Academy of Sciences, 34/5 Vavilov Street, 119334 Moscow, Russia

*To whom correspondence should be addressed. Tel: 7(499)1353092; Fax: 7(499)1354105; Email: sergey.v.razin{at}usa.net

Received April 10, 2008. Revised June 6, 2008. Accepted June 22, 2008.

The spatial configuration of the chicken {alpha}-globin gene domain in erythroid and lymphoid cells was studied by using the Chromosome Conformation Capture (3C) approach. Real-time PCR with TaqMan probes was employed to estimate the frequencies of cross-linking of different restriction fragments within the domain. In differentiated cultured erythroblasts and in 10-day chick embryo erythrocytes expressing ‘adult’ {alpha}A and {alpha}D globin genes the following elements of the domain were found to form an ‘active’ chromatin hub: upstream Major Regulatory Element (MRE), –9 kb upstream DNase I hypersensitive site (DHS), –4 kb upstream CpG island, {alpha}D gene promoter and the downstream enhancer. The {alpha}A gene promoter was not present in the ‘active’ chromatin hub although the level of {alpha}A gene transcription exceeded that of the {alpha}D gene. Formation of the ‘active’ chromatin hub was preceded by the assembly of multiple incomplete hubs containing MRE in combination with either –9 kb DHS or other regulatory elements of the domain. These incomplete chromatin hubs were present in proliferating cultured erythroblasts which did not express globin genes. In lymphoid cells only the interaction between the {alpha}D promoter and the CpG island was detected.


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