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PMID: 15143173 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Role of CTCF binding sites in the Igf2/H19 imprinting control region.

Molecular and cellular biology ·Vol. 24 ·No. 11 ·2004-06-00 ·Pages 4791-800

Szabó PE, Tang SH, Silva FJ, Tsark WM, Mann JR

Abstract

A approximately 2.4-kb imprinting control region (ICR) regulates somatic monoallelic expression of the Igf2 and H19 genes. This is achieved through DNA methylation-dependent chromatin insulator and promoter silencing activities on the maternal and paternal chromosomes, respectively. In somatic cells, the hypomethylated maternally inherited ICR binds the insulator protein CTCF at four sites and blocks activity of the proximal Igf2 promoter by insulating it from its distal enhancers. CTCF binding is thought to play a direct role in inhibiting methylation of the ICR in female germ cells and in somatic cells and, therefore, in establishing and maintaining imprinting of the Igf2/H19 region. Here, we report on the effects of eliminating ICR CTCF binding by severely mutating all four sites in mice. We found that in the female and male germ lines, the mutant ICR remained hypomethylated and hypermethylated, respectively, showing that the CTCF binding sites are dispensable for imprinting establishment. Postfertilization, the maternal mutant ICR acquired methylation, which could be explained by loss of methylation inhibition, which is normally provided by CTCF binding. Adjacent regions in cis-the H19 promoter and gene-also acquired methylation, accompanied by downregulation of H19. This could be the result of a silencing effect of the methylated maternal ICR.

MeSH Terms
Animals Binding Sites CCCTC-Binding Factor DNA Methylation DNA-Binding Proteins/metabolism Gene Expression Regulation/physiology Genomic Imprinting Insulin-Like Growth Factor II/genetics,metabolism Mice Mutation Protein Binding RNA, Long Noncoding RNA, Untranslated/biosynthesis,metabolism Repressor Proteins/metabolism
Chemicals
CCCTC-Binding Factor Ctcf protein, mouse DNA-Binding Proteins H19 long non-coding RNA RNA, Long Noncoding RNA, Untranslated Repressor Proteins Insulin-Like Growth Factor II
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Szabó Piroska E
Division of Biology, Beckman Research Institute of the City of Hope, 1450 East Duarte Rd., Duarte, CA 91010-3011, USA. pszabo@coh.org
Tang Shih-Huey E
Silva Francisco J
Tsark Walter M K
Mann Jeffrey R
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.11.4791-4800.2004
Email
pszabo@coh.org
Published
2004-06-00
Pages
4791-800
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC416431
Subset
IM
Grants
NIGMS NIH HHS · R01 GM064378 · United States
NIGMS NIH HHS · GM064378 · United States
External Links
PubMed source DOI: 10.1128/MCB.24.11.4791-4800.2004 PMC Full Text View journal details
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