CTCF-KDM4A complex correlates with histone modifications that negatively regulate CHD5 gene expression in cancer cell lines

// Lissania Guerra-Calderas 1 , Rodrigo Gonzalez-Barrios 1 , Carlos Cesar Patino 1 , Nicolas Alcaraz 4 , Marisol Salgado-Albarran 1 , David Cantu de Leon 3 , Clementina Castro Hernandez 1, 2 , Yesennia Sanchez-Perez 1 , Hector Aquiles Maldonado-Martinez 5 , Inti A. De la Rosa-Velazquez 6 , Fernanda Vargas-Romero 7 , Luis A. Herrera 1, 2 , Alejandro Garcia-Carranca 1, 2 and Ernesto Soto-Reyes 1 1 Cancer Biomedical Research Unit, Instituto Nacional de Cancerologia (INCan), Mexico City, Mexico 2 Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City, Mexico 3 Clinical Research, Instituto Nacional de Cancerologia (INCan), Mexico City, Mexico 4 The Bioinformatics Centre, Section for RNA and Computational Biology, Department of Biology, University of Copenhagen, Copenhagen, Denmark 5 Department of Surgical Pathology, Instituto Nacional de Cancerologia, Mexico City, Mexico 6 Genomics Lab, Universidad Nacional Autonoma de Mexico, Red de Apoyo a la Investigacion-CIC and Instituto Nacional de Ciencias Medicas y Nutricion "Salvador Zubiran", Mexico City, Mexico 7 Instituto de Fisiologia Celular-Neurociencias, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City, Mexico Correspondence to: Ernesto Soto-Reyes, email: ctcf@ciencias.unam.mx Keywords: KDM4A; CTCF; histone demethylation; H3K36me; CHD5 Received: August 10, 2017      Accepted: February 26, 2018     Published: March 30, 2018 ABSTRACT Histone demethylase KDM4A is involved in H3K9me3 and H3K36me3 demethylation, which are epigenetic modifications associated with gene silencing and RNA Polymerase II elongation, respectively. KDM4A is abnormally expressed in cancer, affecting the expression of multiple targets, such as the CHD5 gene. This enzyme localizes at the first intron of CHD5 , and the dissociation of KDM4A increases gene expression. In vitro assays showed that KDM4A-mediated demethylation is enhanced in the presence of CTCF, suggesting that CTCF could increase its enzymatic activity in vivo, however the specific mechanism by which CTCF and KDM4A might be involved in the CHD5 gene repression is poorly understood. Here, we show that CTCF and KDM4A form a protein complex, which is recruited into the first intron of CHD5 . This is related to a decrease in H3K36me3/2 histone marks and is associated with its transcriptional downregulation. Depletion of CTCF or KDM4A by siRNA, triggered the reactivation of CHD5 expression, suggesting that both proteins are involved in the negative regulation of this gene. Furthermore, the knockout of KDM4A restored the CHD5 expression and H3K36me3 and H3K36me2 histone marks. Such mechanism acts independently of CHD5 promoter DNA methylation. Our findings support a novel mechanism of epigenetic repression at the gene body that does not involve promoter silencing.