Functional association of CTCF with the insulator upstream of the H19 gene is parent of origin-specific and methylation-sensitive

Curr Biol. 2000 Jul 13;10(14):853-6. doi: 10.1016/s0960-9822(00)00597-2.


In mammals, a subset of genes inherit gametic marks that establish parent of origin-dependent expression patterns in the soma ([1] and references therein). The currently most extensively studied examples of this phenomenon, termed genomic imprinting, are the physically linked Igf2 (insulin-like growth factor II) and H19 genes, which are expressed mono-allelically from opposite parental alleles [1] [2]. The repressed status of the maternal Igf2 allele is due to cis elements that prevent the H19 enhancers [3] from accessing the Igf2 promoters on the maternal chromosome [4] [5]. A differentially methylated domain (DMD) in the 5' flank of H19 is maintained paternally methylated and maternally unmethylated [6] [7]. We show here by gel-shift and chromatin immunopurification analyses that binding of the highly conserved multivalent factor CTCF ([8] [9] and references therein) to the H19 DMD is methylation-sensitive and parent of origin-dependent. Selectively mutating CTCF-contacting nucleotides, which were identified by methylation interference within the extended binding sites initially revealed by nuclease footprinting, abrogated the H19 DMD enhancer-blocking property. These observations suggest that molecular mechanisms of genomic imprinting may use an unusual ability of CTCF to interact with a diverse spectrum of variant target sites, some of which include CpGs that are responsible for methylation-sensitive CTCF binding in vitro and in vivo.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites / genetics
  • CCCTC-Binding Factor
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Methylation
  • DNA-Binding Proteins / metabolism*
  • Enhancer Elements, Genetic
  • Female
  • Insulin-Like Growth Factor II / genetics
  • Male
  • Mice
  • Molecular Sequence Data
  • Muscle Proteins / genetics*
  • Protein Binding
  • RNA, Long Noncoding
  • RNA, Untranslated*
  • Repressor Proteins*
  • Transcription Factors / metabolism*
  • Zinc Fingers


  • CCCTC-Binding Factor
  • Ctcf protein, mouse
  • DNA-Binding Proteins
  • H19 long non-coding RNA
  • Muscle Proteins
  • RNA, Long Noncoding
  • RNA, Untranslated
  • Repressor Proteins
  • Transcription Factors
  • Insulin-Like Growth Factor II
  • DNA