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The transcriptional regulator CTCF is involved in the control of ribosomal DNA

AuthorsDelgado, M. Dolores ; Rosa-Garrido, Manuel ; Batlle-López, Ana
Issue Date2010
CitationEACR 21 (2010)
Abstract[Background]: CTCF is a ubiquitously expressed transcriptional regulator that binds at multiple sites throughout the genome. CTCF is a putative oncosuppressor with several funetions such as regulalion of cell proliferation, differen tiation, apoptosis, enhaneer-blocking activity and control of imprinted genes. Our previous studies on CTCF subnuclear localization and in situ runon assays revealed that CTCF was associated with several components of the nucleolus and influenced nucleolar transcription (Torrano et al JCS 2006). In this work we asked whether CTCF could have a direct role in the regulation of ribosomal genes transcription. [Material and Methods]: CTCF binding sites in the human rDNAwere searched with a CTCF Binding Sites Data Base (Bao et al NAR 2008). In vitro binding of CTCF and UBF tQ rDNA was studied by Electrophoretic Mobility Shift Assays (EMSA). Oecupancy of CTCF, UBF and modified hislones to ribosomal DNA was analyzed by Chromatin Immunoprecipitation (ChiP). [Results]: We found several putative CTCF binding sites to human rDNA. EMSA analysis revealed CTCF binding to two sites mapped in the intergenic spacer region S' upstream of the ribosomal gene promoter. CTCF binding to both sites was methylalion-sensitive. ChiP analysis showed in vivo occupancy of CTCF, as well as its paraloge CTCFL (BORI S), to the rDNA. The transcription factor of the RNA polymerase I UBF plays a critical role on the regulation of ribosomal genes transcription. By serial-ChiP assays we found interaction of both CTCF and UBF at both rONA sites. Variant histone H2AZ and marks of active chromatin were present at CTCF binding sites on ribosomal genes. [Conclusions]: We have identified two CTCF binding sites upstream of the human rDNA promoter. CTCF in vitro binding and in vivo occupancy at both rONA sites as well as interaclion of CTCF and·UBF at rONA was demonstrated. Our results showed a novel funetion of CTCF on the ribosomal biogenesis regulation, a critical process linked to the control of ce1l prolifera!ion and differentiation.
DescriptionTrabajo presentado al "21th Meeting of the European Association for Cancer Research" celebrado en Oslo (Noruega) del 26 al 29 de junio de 2010.-- et al.
Appears in Collections:(IBBTEC) Comunicaciones congresos
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