ChromID identifies the protein interactome at chromatin marks.
Nat Biotechnol
; 38(6): 728-736, 2020 06.
Article
em En
| MEDLINE
| ID: mdl-32123383
ABSTRACT
Chromatin modifications regulate genome function by recruiting proteins to the genome. However, the protein composition at distinct chromatin modifications has yet to be fully characterized. In this study, we used natural protein domains as modular building blocks to develop engineered chromatin readers (eCRs) selective for DNA methylation and histone tri-methylation at H3K4, H3K9 and H3K27 residues. We first demonstrated their utility as selective chromatin binders in living cells by stably expressing eCRs in mouse embryonic stem cells and measuring their subnuclear localization, genomic distribution and histone-modification-binding preference. By fusing eCRs to the biotin ligase BASU, we established ChromID, a method for identifying the chromatin-dependent protein interactome on the basis of proximity biotinylation, and applied it to distinct chromatin modifications in mouse stem cells. Using a synthetic dual-modification reader, we also uncovered the protein composition at bivalently modified promoters marked by H3K4me3 and H3K27me3. These results highlight the ability of ChromID to obtain a detailed view of protein interaction networks on chromatin.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Cromatina
/
Histonas
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Mapeamento de Interação de Proteínas
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Proteômica
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Mapas de Interação de Proteínas
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article