Analysis of protein-DNA interactions in chromatin by UV induced cross-linking and mass spectrometry.

ABSTRACT: Protein-DNA interactions are key to the functionality and stability of the genome. Identification and mapping of protein-DNA interaction interfaces and sites is crucial for understanding DNA-dependent processes. Here, we present a workflow that allows mass spectrometric (MS) identification of proteins in direct contact with DNA in reconstituted and native chromatin after cross-linking by ultraviolet (UV) light. Our approach enables the determination of contact interfaces at amino-acid level. With the example of chromatin-associated protein SCML2 we show that our technique allows differentiation of nucleosome-binding interfaces in distinct states. By UV cross-linking of isolated nuclei we determined the cross-linking sites of several factors including chromatin-modifying enzymes, demonstrat

SUBMITTER: Stutzer A 

PROVIDER: S-EPMC7567871 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature