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Autor(en): Mauser, Rebekka
Kungulovski, Goran
Keup, Corinna
Reinhardt, Richard
Jeltsch, Albert
Titel: Application of dual reading domains as novel reagents in chromatin biology reveals a new H3K9me3 and H3K36me2/3 bivalent chromatin state
Erscheinungsdatum: 2017
Dokumentart: Zeitschriftenartikel
Erschienen in: Epigenetics & chromatin 10 (2017), No. 45
URI: http://elib.uni-stuttgart.de/handle/11682/9257
http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-92571
http://dx.doi.org/10.18419/opus-9240
ISSN: 1756-8935
Zusammenfassung: Histone post-translational modifications (PTMs) play central roles in chromatin-templated processes. Combinations of two or more histone PTMs form unique interfaces for readout and recruitment of chromatin-interacting complexes, but the genome-wide mapping of co-existing histone PTMs remains an experimentally difficult task. We introduce here a novel type of affinity reagents consisting of two fused recombinant histone modification interacting domains (HiMID) for direct detection of doubly modified chromatin. To develop the method, we fused the MPP8 Chromodomain and DNMT3A PWWP domain which have a binding specificity for H3K9me3 and H3K36me2/3, respectively. We validate the novel reagent biochemically and in ChIP applications and show its specific interaction with H3K9me3-H3K36me2/3 doubly modified chromatin. Modification specificity was confirmed using mutant double-HiMIDs with inactivated methyllysine binding pockets. Using this novel tool, we mapped co-existing H3K9me3-H3K36me2/3 marks in human cells by chromatin interaction domain precipitation (CIDOP). CIDOP-seq data were validated by qPCR, sequential CIDOP/ChIP and by comparison with CIDOP- and ChIP-seq data obtained with single modification readers and antibodies. The genome-wide distribution of H3K9me3-H3K36me2/3 indicates that it represents a novel bivalent chromatin state, which is enriched in weakly transcribed chromatin segments and at ZNF274 and SetDB1 binding sites.
Enthalten in den Sammlungen:03 Fakultät Chemie

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