Unlike some reality show marriages that crack under the media spotlight, the union of automation and big data is still going strong, especially when it comes to cataloging histone post-translational modifications (PTM). Researchers from Princeton University show off a new systematic approach for figuring out just what all those histone PTM combinations are up to in chromatin. The novel technology is based on streamlined semisynthesis of DNA-barcoded nucleosome libraries with distinct PTM combos. Here’s what that all means:
- DNA-barcoded nucleosome libraries can be generated to create your histone PTM pattern of interest.
- These libraries can be treated with induvidual purified chromatin effectors or any combination of these readers, writers, and erasers.
- The nucleosomes containing the custom histone patterns and effectors of interest are then subjected to ChIP and multiplexed DNA-barcode sequencing.
Ultimately, this ultra sensitive new workflow allows for the collection of thousands of biochemical data points that decrypt the controls of chromatin. It allows for the understanding of how the nuclear proteome interacts and binds with a single PTM or complex combos. Not only that, but it can also be used to look deeper into the histone code to see how the binding preferences of nuclear factors affect PTM patterns via cross-talk when in combination.
This technique offers a versatile new platform that greatly accelerates biochemical investigations into chromatin recognition and signalling at the level of single PTMs, PTM combos, as well as the effect of nuclear factors on PTM patterns.
ChIP away at the histone code combo complexities in Nature Methods, July 2014