Accelerated chromatin biochemistry using dnA-barcoded nucleosome libraries

Uyen T.T. Nguyen, Lenka Bittova, Manuel M. Müller, Beat Fierz, Yael David, Brian Houck-Loomis, Vanessa Feng, Geoffrey P. Dann, Tom W. Muir

Research output: Contribution to journalArticle

82 Scopus citations

Abstract

Elucidating the molecular details of how chromatin-associated factors deposit, remove and recognize histone post-translational modifcation (Ptm) signatures remains a daunting task in the epigenetics feld. We introduce a versatile platform that greatly accelerates biochemical investigations into chromatin recognition and signaling. this technology is based on the streamlined semisynthesis of dnA-barcoded nucleosome libraries with distinct combinations of Ptms. chromatin immunoprecipitation of these libraries, once they have been treated with purifed chromatin effectors or the combined chromatin recognizing and modifying activities of the nuclear proteome, is followed by multiplexed dnA-barcode sequencing. this ultrasensitive workfow allowed us to collect thousands of biochemical data points revealing the binding preferences of various nuclear factors for Ptm patterns and how preexisting Ptms, alone or synergistically, affect further Ptm deposition via cross-talk mechanisms. We anticipate that the high throughput and sensitivity of the technology will help accelerate the decryption of the diverse molecular controls that operate at the level of chromatin.

Original languageEnglish (US)
Pages (from-to)834-840
Number of pages7
JournalNature Methods
Volume11
Issue number8
DOIs
StatePublished - Aug 2014

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Nguyen, U. T. T., Bittova, L., Müller, M. M., Fierz, B., David, Y., Houck-Loomis, B., Feng, V., Dann, G. P., & Muir, T. W. (2014). Accelerated chromatin biochemistry using dnA-barcoded nucleosome libraries. Nature Methods, 11(8), 834-840. https://doi.org/10.1038/nMeth.3022