Dynamic CTCF binding directly mediates interactions among cis-regulatory elements essential for hematopoiesis

Blood. 2021 Mar 11;137(10):1327-1339. doi: 10.1182/blood.2020005780.

Abstract

While constitutive CCCTC-binding factor (CTCF)-binding sites are needed to maintain relatively invariant chromatin structures, such as topologically associating domains, the precise roles of CTCF to control cell-type-specific transcriptional regulation remain poorly explored. We examined CTCF occupancy in different types of primary blood cells derived from the same donor to elucidate a new role for CTCF in gene regulation during blood cell development. We identified dynamic, cell-type-specific binding sites for CTCF that colocalize with lineage-specific transcription factors. These dynamic sites are enriched for single-nucleotide polymorphisms that are associated with blood cell traits in different linages, and they coincide with the key regulatory elements governing hematopoiesis. CRISPR-Cas9-based perturbation experiments demonstrated that these dynamic CTCF-binding sites play a critical role in red blood cell development. Furthermore, precise deletion of CTCF-binding motifs in dynamic sites abolished interactions of erythroid genes, such as RBM38, with their associated enhancers and led to abnormal erythropoiesis. These results suggest a novel, cell-type-specific function for CTCF in which it may serve to facilitate interaction of distal regulatory emblements with target promoters. Our study of the dynamic, cell-type-specific binding and function of CTCF provides new insights into transcriptional regulation during hematopoiesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Binding Sites
  • CCCTC-Binding Factor / metabolism*
  • Cell Line
  • Cells, Cultured
  • Enhancer Elements, Genetic
  • Erythroid Cells / cytology
  • Erythroid Cells / metabolism
  • Erythropoiesis*
  • Humans
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • Regulatory Elements, Transcriptional*
  • Transcriptional Activation

Substances

  • CCCTC-Binding Factor
  • CTCF protein, human
  • RBM38 protein, human
  • RNA-Binding Proteins