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Molecular and Cellular Biology, March 2006, p. 1589-1597, Vol. 26, No. 5
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.5.1589-1597.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A CTCF-Dependent Silencer Located in the Differentially Methylated Area May Regulate Expression of a Housekeeping Gene Overlapping a Tissue-Specific Gene Domain

Denis Klochkov,1 Héctor Rincón-Arano,2 Elena S. Ioudinkova,1 Viviana Valadez-Graham,2 Alexey Gavrilov,1,3 Félix Recillas-Targa,2 and Sergey V. Razin1,3*

Laboratory of Structural-Functional Organization of Chromosomes, Institute of Gene Biology of the Russian Academy of Sciences, 34/5 Vavilov Street, 117334 Moscow, Russia,1 Instituto de Fisiología Celular, Departamento de Genética Molecular, Universidad Nacional Autónoma de México, Apartado Postal 70-242, México D.F. 045019, México,2 Department of Molecular Biology, Moscow State University, 119899 Moscow, Russia3

Received 1 August 2005/ Returned for modification 18 September 2005/ Accepted 4 December 2005

The tissue-specific chicken {alpha}-globin gene domain represents one of the paradigms, in terms of its constitutively open chromatin conformation and the location of several regulatory elements within the neighboring housekeeping gene. Here, we show that an 0.2-kb DNA fragment located ~4 kb upstream to the chicken {alpha}-globin gene cluster contains a binding site for the multifunctional protein factor CTCF and possesses silencer activity which depends on CTCF binding, as demonstrated by site-directed mutagenesis of the CTCF recognition sequence. CTCF was found to be associated with this recognition site in erythroid cells but not in lymphoid cells where the site is methylated. A functional promoter directing the transcription of the apparently housekeeping ggPRX gene was found 120 bp from the CTCF-dependent silencer. The data are discussed in terms of the hypothesis that the CTCF-dependent silencer stabilizes the level of ggPRX gene transcription in erythroid cells where the promoter of this gene may be influenced by positive cis-regulatory signals activating {alpha}-globin gene transcription.

* Corresponding author. Mailing address: Laboratory of Structural-Functional Organization of Chromosomes, Institute of Gene Biology of the Russian Academy of Sciences, 34/5 Vavilov Street, 117334 Moscow, Russia. Phone: 7-095-135-3092. Fax: 7-095-135-4105. E-mail: sergey.v.razin{at}usa.net.

Molecular and Cellular Biology, March 2006, p. 1589-1597, Vol. 26, No. 5
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.5.1589-1597.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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