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Measuring Chromatin Interaction Dynamics on the Second Time Scale at Single-Copy Genes

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Science  03 Oct 2013:
1242369
DOI: 10.1126/science.1242369

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Abstract

The chromatin immunoprecipitation (ChIP) assay is widely used to capture interactions between chromatin and regulatory proteins, but it is unknown how stable most native interactions are. Although live cell imaging suggests short-lived interactions at tandem gene arrays, current methods cannot measure rapid binding dynamics at single-copy genes. Here, we show using a modified ChIP assay with sub-second temporal resolution that the time dependence of formaldehyde cross-linking can be used to extract in vivo on- and off-rates for site-specific chromatin interactions varying over a ~100-fold dynamic range. Using the method, we show that a regulatory process can shift weakly bound TATA-binding protein to stable promoter interactions, thereby facilitating transcription complex formation. This assay provides an approach for systematic, quantitative analyses of chromatin binding dynamics in vivo.

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