Rapid degradation of DHX36 revealing its transcriptional role by interacting with G-quadruplex
More evident supports G-quadruplex are involved in transcription. Recent research revealed that DHX36 preferentially resolves G-quadruplex (G4) structure over the canonical G4, raising the possibility that DHX36 could bind and resolve G4 structures to regulate gene transcription. However, the hypothesis has yet been validated systematically in vivo.In this study, we investigated the role of DHX36 interacting with G4s in transcription. Firstly, we performed CUT&TAG to map the binding sites in chromatin of MCF7. Next, we utilized nascent RNA-seq and AID protein degradation system to identify the direct gene targets by transcriptional regulation of DHX36 and found these sites are enriched with G4 structure. We picked three G4 sequences in oncogene from these sites and found DHX36 binds and resolves these G4 structures in vitro, suggesting DHX36 may be recruited to these promoter G4 sites and regulate gene transcription by resolving G4 structure. Overall design: Cleavage Under Target & Tagmentation (CUT&TAG) for DHX36 in MCF7 cells, and nascent RNA-seq acrylonitrile-mediated uridine-to-cytidine conversion (AMUC-seq) with or without depletion of DHX36 by Auxin Inducible Degron (AID) assay.