Spatiotemporal and global profiling of DNA-protein interactions enables discovery of low-affinity transcription factors

ABSTRACT: Precise dissection of DNA-protein interactions is essential for elucidating recognition basis, dynamics, and gene regulation mechanism. However, global profiling of weak and dynamic DNA-protein interactions remains a long-standing challenge. Here, we establish light-induced lysine (K) enabled crosslinking (LIKE-XL) strategy for spatiotemporal and global profiling of DNA-protein interactions. Harnessing the unique abilities to capture weak and transient DNA-protein interactions, we demonstrate that LIKE-XL enables discovery of low-affinity transcription factors (TFs)-DNA interactions via sequence-specific DNA baits, determining the binding sites for TFs previously unachieved. More importantly, we successfully decipher the dynamics of TF subproteome in response to drug treatment in time-resolved manner, and find new downstream target TFs from drug perturbations, providing insights into their dynamic transcriptional networks. The LIKE-XL offers a novel and complementary method to expand the DNA-protein profiling toolbox and map accurate DNA-protein interactomes previously inaccessible via noncovalent strategies, for better understanding of protein functions in health and disease.

ORGANISM(S): Homo Sapiens

SUBMITTER: Minjia Tan  

PROVIDER: PXD039229 | iProX | Tue Jan 03 00:00:00 GMT 2023

REPOSITORIES: iProX