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  1. Article: Critical DNA binding interactions of the insulator protein CTCF: a small number of zinc fingers mediate strong binding, and a single finger-DNA interaction controls binding at imprinted loci.

    Renda, Mario / Baglivo, Ilaria / Burgess-Beusse, Bonnie / Esposito, Sabrina / Fattorusso, Roberto / Felsenfeld, Gary / Pedone, Paolo V

    The Journal of biological chemistry

    2007  Volume 282, Issue 46, Page(s) 33336–33345

    Abstract: The DNA-binding protein CTCF (CCCTC binding factor) mediates enhancer blocking insulation at sites throughout the genome and plays an important role in regulating allele-specific expression at the Igf2/H19 locus and at other imprinted loci. Evidence is ... ...

    Abstract The DNA-binding protein CTCF (CCCTC binding factor) mediates enhancer blocking insulation at sites throughout the genome and plays an important role in regulating allele-specific expression at the Igf2/H19 locus and at other imprinted loci. Evidence is also accumulating that CTCF is involved in large scale organization of genomic chromatin. Although CTCF has 11 zinc fingers, we show here that only 4 of these are essential to strong binding and that they recognize a core 12-bp DNA sequence common to most CTCF sites. By deleting individual fingers and mutating individual sites, we determined the orientation of binding. Furthermore, we were able to identify the specific finger and its point of DNA interaction that are responsible for the loss of CTCF binding when CpG residues are methylated in the imprinted Igf2/H19 locus. This single interaction appears to be critical for allele-specific binding and insulation by CTCF.
    MeSH term(s) Base Sequence ; Binding Sites ; CCCTC-Binding Factor ; CpG Islands ; DNA/chemistry ; DNA Methylation ; DNA-Binding Proteins/chemistry ; DNA-Binding Proteins/physiology ; Enhancer Elements, Genetic ; Genomic Imprinting ; Humans ; Molecular Sequence Data ; Protein Binding ; Protein Structure, Tertiary ; Repressor Proteins/chemistry ; Repressor Proteins/physiology ; Transcription Factors/metabolism ; Zinc Fingers
    Chemical Substances CCCTC-Binding Factor ; CTCF protein, human ; DNA-Binding Proteins ; Repressor Proteins ; Transcription Factors ; DNA (9007-49-2)
    Language English
    Publishing date 2007-09-07
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1074/jbc.M706213200
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Uc I Zs.108: Show issues Location:
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  2. Article: Critical DNA Binding Interactions of the Insulator Protein CTCF: A SMALL NUMBER OF ZINC FINGERS MEDIATE STRONG BINDING, AND A SINGLE FINGER-DNA INTERACTION CONTROLS BINDING AT IMPRINTED LOCI

    Renda, Mario / Baglivo, Ilaria / Burgess-Beusse, Bonnie / Esposito, Sabrina / Fattorusso, Roberto / Felsenfeld, Gary / Pedone, Paolo V

    Journal of biological chemistry. 2007 Nov. 16, v. 282, no. 46

    2007  

    Abstract: The DNA-binding protein CTCF (CCCTC binding factor) mediates enhancer blocking insulation at sites throughout the genome and plays an important role in regulating allele-specific expression at the Igf2/H19 locus and at other imprinted loci. Evidence is ... ...

    Abstract The DNA-binding protein CTCF (CCCTC binding factor) mediates enhancer blocking insulation at sites throughout the genome and plays an important role in regulating allele-specific expression at the Igf2/H19 locus and at other imprinted loci. Evidence is also accumulating that CTCF is involved in large scale organization of genomic chromatin. Although CTCF has 11 zinc fingers, we show here that only 4 of these are essential to strong binding and that they recognize a core 12-bp DNA sequence common to most CTCF sites. By deleting individual fingers and mutating individual sites, we determined the orientation of binding. Furthermore, we were able to identify the specific finger and its point of DNA interaction that are responsible for the loss of CTCF binding when CpG residues are methylated in the imprinted Igf2/H19 locus. This single interaction appears to be critical for allele-specific binding and insulation by CTCF.
    Language English
    Dates of publication 2007-1116
    Size p. 33336-33345.
    Publishing place American Society for Biochemistry and Molecular Biology
    Document type Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    Database NAL-Catalogue (AGRICOLA)

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  3. Article: Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities.

    Recillas-Targa, Félix / Pikaart, Michael J / Burgess-Beusse, Bonnie / Bell, Adam C / Litt, Michael D / West, Adam G / Gaszner, Miklos / Felsenfeld, Gary

    Proceedings of the National Academy of Sciences of the United States of America

    2002  Volume 99, Issue 10, Page(s) 6883–6888

    Abstract: The 1.2-kb DNA sequence element (5'HS4) at the 5' end of the chicken beta-globin locus has the two defining properties of an insulator: it prevents an "external" enhancer from acting on a promoter when placed between them ("enhancer blocking") and acts ... ...

    Abstract The 1.2-kb DNA sequence element (5'HS4) at the 5' end of the chicken beta-globin locus has the two defining properties of an insulator: it prevents an "external" enhancer from acting on a promoter when placed between them ("enhancer blocking") and acts as a barrier to chromosomal position effect (CPE) when it surrounds a stably integrated reporter. We previously reported that a single CTCF-binding site in 5'HS4 is necessary and sufficient for enhancer blocking. We show here that a 250-bp "core" element from within 5'HS4 is sufficient to confer protection against silencing of transgenes caused by CPE. Further dissection of the core reveals that 5'HS4 is a compound element in which it is possible to separate enhancer blocking and barrier activities. We demonstrate that full protection against CPE is conferred by mutant 5'HS4 sequences from which the CTCF-binding site has been deleted. In contrast, mutations of four other protein binding sites within 5'HS4 result in varying reductions in the ability to protect against CPE. We find that binding sites for CTCF are neither necessary nor sufficient for protection against CPE. Comparison of the properties of 5'HS4 with those of other CTCF-binding enhancer-blocking elements suggests that CPE protection is associated with maintenance of a high level of histone acetylation near the insulator, conferred by insulator binding-proteins other than CTCF.
    MeSH term(s) Animals ; CCCTC-Binding Factor ; Cell Line, Transformed ; Chickens ; DNA-Binding Proteins/metabolism ; Enhancer Elements, Genetic ; Globins/genetics ; Repressor Proteins ; Transcription Factors/metabolism
    Chemical Substances CCCTC-Binding Factor ; DNA-Binding Proteins ; Repressor Proteins ; Transcription Factors ; Globins (9004-22-2)
    Language English
    Publishing date 2002-05-14
    Publishing country United States
    Document type Journal Article
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.102179399
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
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  4. Article: The insulation of genes from external enhancers and silencing chromatin.

    Burgess-Beusse, Bonnie / Farrell, Catherine / Gaszner, Miklos / Litt, Michael / Mutskov, Vesco / Recillas-Targa, Felix / Simpson, Melanie / West, Adam / Felsenfeld, Gary

    Proceedings of the National Academy of Sciences of the United States of America

    2002  Volume 99 Suppl 4, Page(s) 16433–16437

    Abstract: Insulators are DNA sequence elements that can serve in some cases as barriers to protect a gene against the encroachment of adjacent inactive condensed chromatin. Some insulators also can act as blocking elements to protect against the activating ... ...

    Abstract Insulators are DNA sequence elements that can serve in some cases as barriers to protect a gene against the encroachment of adjacent inactive condensed chromatin. Some insulators also can act as blocking elements to protect against the activating influence of distal enhancers associated with other genes. Although most of the insulators identified so far derive from Drosophila, they also are found in vertebrates. An insulator at the 5' end of the chicken beta-globin locus marks a boundary between an open chromatin domain and a region of constitutively condensed chromatin. Detailed analysis of this element shows that it possesses both enhancer blocking activity and the ability to screen reporter genes against position effects. Enhancer blocking is associated with binding of the protein CTCF; sites that bind CTCF are found at other critical points in the genome. Protection against position effects involves other properties that appear to be associated with control of histone acetylation and methylation. Insulators thus are complex elements that can help to preserve the independent function of genes embedded in a genome in which they are surrounded by regulatory signals they must ignore.
    MeSH term(s) Animals ; Chromatin/genetics ; Enhancer Elements, Genetic ; Gene Silencing ; Humans
    Chemical Substances Chromatin
    Language English
    Publishing date 2002-12-10
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.162342499
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
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