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Mutational Analysis of the Poly(ADP-Ribosyl)ation Sites of the Transcription Factor CTCF Provides an Insight into the Mechanism of Its Regulation by Poly(ADP-Ribosyl)ation

Farrar, Dawn and Rai, Sushma and Chernukhin, Igor and Jagodic, Maja and Ito, Yoko and Yammine, Samer and Ohlsson, Rolf and Murrell, Adele and Klenova, Elena (2010) 'Mutational Analysis of the Poly(ADP-Ribosyl)ation Sites of the Transcription Factor CTCF Provides an Insight into the Mechanism of Its Regulation by Poly(ADP-Ribosyl)ation.' Molecular and Cellular Biology, 30 (5). pp. 1199-1216. ISSN 0270-7306

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Abstract

<jats:title>ABSTRACT</jats:title> <jats:p> Poly(ADP-ribosyl)ation of the conserved multifunctional transcription factor CTCF was previously identified as important to maintain CTCF insulator and chromatin barrier functions. However, the molecular mechanism of this regulation and also the necessity of this modification for other CTCF functions remain unknown. In this study, we identified potential sites of poly(ADP-ribosyl)ation within the N-terminal domain of CTCF and generated a mutant deficient in poly(ADP-ribosyl)ation. Using this CTCF mutant, we demonstrated the requirement of poly(ADP-ribosyl)ation for optimal CTCF function in transcriptional activation of the p <jats:italic>19ARF</jats:italic> promoter and inhibition of cell proliferation. By using a newly generated isogenic insulator reporter cell line, the CTCF insulator function at the mouse <jats:italic>Igf2-H19</jats:italic> imprinting control region (ICR) was found to be compromised by the CTCF mutation. The association and simultaneous presence of PARP-1 and CTCF at the ICR, confirmed by single and serial chromatin immunoprecipitation assays, were found to be independent of CTCF poly(ADP-ribosyl)ation. These results suggest a model of CTCF regulation by poly(ADP-ribosyl)ation whereby CTCF and PARP-1 form functional complexes at sites along the DNA, producing a dynamic reversible modification of CTCF. By using bioinformatics tools, numerous sites of CTCF and PARP-1 colocalization were demonstrated, suggesting that such regulation of CTCF may take place at the genome level. </jats:p>

Item Type: Article
Uncontrolled Keywords: Cell Line; Hela Cells; Hybrid Cells; Animals; Humans; Mice; Poly(ADP-ribose) Polymerases; Poly Adenosine Diphosphate Ribose; Insulin-Like Growth Factor II; Recombinant Proteins; Repressor Proteins; RNA, Untranslated; DNA Primers; Transfection; Amino Acid Substitution; Mutagenesis, Site-Directed; Cell Proliferation; Genomic Imprinting; Binding Sites; Amino Acid Sequence; Base Sequence; Mutation; Molecular Sequence Data; RNA, Long Noncoding; Poly (ADP-Ribose) Polymerase-1; CCCTC-Binding Factor
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Science and Health
Faculty of Science and Health > Life Sciences, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 07 Sep 2011 13:27
Last Modified: 14 Mar 2022 21:48
URI: http://repository.essex.ac.uk/id/eprint/78

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