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Novel CTCF binding at a site in exon1A of BCL6 is associated with active histone marks and a transcriptionally active locus

Oncogene volume 34, pages 246–256 (2015)Cite this article

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Abstract

BCL6 is a zinc-finger transcriptional repressor, which is highly expressed in germinal centre B-cells and is essential for germinal centre formation and T-dependent antibody responses. Constitutive BCL6 expression is sufficient to produce lymphomas in mice. Deregulated expression of BCL6 due to chromosomal rearrangements, mutations of a negative autoregulatory site in the BCL6 promoter region and aberrant post-translational modifications have been detected in a number of human lymphomas. Tight lineage and temporal regulation of BCL6 is, therefore, required for normal immunity, and abnormal regulation occurs in lymphomas. CCCTC-binding factor (CTCF) is a multi-functional chromatin regulator, which has recently been shown to bind in a methylation-sensitive manner to sites within the BCL6 first intron. We demonstrate a novel CTCF-binding site in BCL6 exon1A within a potential CpG island, which is unmethylated both in cell lines and in primary lymphoma samples. CTCF binding, which was found in BCL6-expressing cell lines, correlated with the presence of histone variant H2A.Z and active histone marks, suggesting that CTCF induces chromatin modification at a transcriptionally active BCL6 locus. CTCF binding to exon1A was required to maintain BCL6 expression in germinal centre cells by avoiding BCL6-negative autoregulation. Silencing of CTCF in BCL6-expressing cells reduced BCL6 mRNA and protein expression, which is sufficient to induce B-cell terminal differentiation toward plasma cells. Moreover, lack of CTCF binding to exon1A shifts the BCL6 local chromatin from an active to a repressive state. This work demonstrates that, in contexts in which BCL6 is expressed, CTCF binding to BCL6 exon1A associates with epigenetic modifications indicative of transcriptionally open chromatin.

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Acknowledgements

We thank Elena Klenova and Rainer Renkawitz for CTCF expression vectors, Miguel Angel Piris and Noemi Nagy for cell lines and Javier León for useful comments on the manuscript. We are grateful to the Department of Hematology of Hospital U. Marques de Valdecilla (Santander) for help with patient recruitment. Funding for this work was provided by Instituto de Salud Carlos III grants PI11/00397 (Fondo Investigaciones Sanitarias) and RETIC (Red Tematica de Investigacion Cooperativa en Cancer) RD06/0020/0017 and RD12/0036/0033. AB was supported by a López Albo fellowship (IFIMAV, Spain), MRG by a PhD fellowship from the University of Cantabria and VT by the Lady Tata Memorial Trust. Part of this work is in AB’s PhD dissertation (split PhD Programme between Imperial College London and the University of Cantabria).

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Author notes

  1. M Rosa-Garrido

    Present address: 4Current address: Department of Anesthesiology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.,

  2. V Torrano

    Present address: 5Current address: Proteomics Unit, CIC bioGUNE, Bizkaia Technology Park, 801 Building, 48160 Derio, Spain.,

Authors and Affiliations

  1. Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC) and Departamento de Biología Molecular, Universidad de Cantabria, CSIC, SODERCAN, Santander, Spain

    A Batlle-López, M G Cortiguera, M Rosa-Garrido, R Blanco, V Torrano & M D Delgado

  2. Servicio de Hematología, Hospital U. Marqués de Valdecilla, and IFIMAV-FMV, Santander, Spain

    A Batlle-López, M G Cortiguera & E del Cerro

  3. Department of Cancer Studies and Molecular Medicine and MRC Toxicology Unit, University of Leicester, Leicester, UK

    S D Wagner

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  1. A Batlle-López
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Correspondence to S D Wagner or M D Delgado.

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Batlle-López, A., Cortiguera, M., Rosa-Garrido, M. et al. Novel CTCF binding at a site in exon1A of BCL6 is associated with active histone marks and a transcriptionally active locus. Oncogene 34, 246–256 (2015). https://doi.org/10.1038/onc.2013.535

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