DDX21 mediates co-transcriptional RNA m6A modification to promote transcription termination and genome stability

Abstract

N6-methyladenosine (m⁶A) is a crucial RNA modification that regulates diverse biological processes in human cells, but its co-transcriptional deposition and functions remain poorly understood. Here, we identified the RNA helicase DDX21 with a previously unrecognized role in directing m⁶A modification on nascent RNA for co-transcriptional regulation. DDX21 interacts with METTL3 for co-recruitment to chromatin through its recognition of R-loops, which can be formed co-transcriptionally as nascent transcripts hybridize onto the template DNA strand. Moreover, DDX21's helicase activity is needed for METTL3-mediated m⁶A deposition onto nascent RNA following recruitment. At transcription termination regions, this nexus of actions promotes XRN2-mediated termination of RNAPII transcription. Disruption of any of these steps, including the loss of DDX21, METTL3, or their enzymatic activities, leads to defective termination that can induce DNA damage. Therefore, we propose that the R-loop-DDX21-METTL3 nexus forges the missing link for co-transcriptional modification of m⁶A, coordinating transcription termination and genome stability.

Keywords: DDX21; DNA damage; METTL3; R-loops; RNA m(6)A methylation; XRN2; co-transcriptional regulation; genome stability; transcription termination; transcription-replication collisions.