期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 105, 期 11, 页码 4093-4098出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0710515105
关键词
electron transfer dissociation; epigenetics; posttranslational modification; histone code; pluripotency
资金
- NHGRI NIH HHS [5T32HG002706] Funding Source: Medline
- NIGMS NIH HHS [1R01GM080148, R01 GM080148-02, 5T32GM08349, R01 GM080148-01A1, R01 GM080148, T32 GM008349, R01 GM080148-03] Funding Source: Medline
Epigenetic regulation through chromatin is thought to play a critical role in the establishment and maintenance of pluripotency. Traditionally, antibody-based technologies were used to probe for specific posttranslational modifications (PTMs) present on histone tails, but these methods do not generally reveal the presence of multiple modifications on a single-histone tail (combinatorial codes). Here, we describe technology for the discovery and quantification of histone combinatorial codes that is based on chromatography and mass spectrometry. We applied this methodology to decipher 74 discrete combinatorial codes on the tail of histone H4 from human embryonic stem (ES) cells. Finally, we quantified the abundances of these codes as human ES cells undergo differentiation to reveal striking changes in methylation and acetylation patterns. For example, H4R3 methylation was observed only in the presence of H4K20 dimethylation; such context-specific patterning exemplifies the power of this technique.
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