Alphafold2 modeling of KCTD10 interactions with RNAPII machinery

During DNA replication, the replisome must remove barriers and roadblocks including the transcription machinery. Transcription-replication conflicts (TRCs) occur when there are collisions between the replisome and transcription machinery, and are increasingly recognized as an important source of mammalian genome instability. How cells facilitate replisome bypass at sites of TRCs is incompletely understood. Here, we investigated the role of the CUL3-KCTD10 E3 ligase as a sensor for TRCs. We found that the substrate adaptor KCTD10 interacts with both the replisome and transcription machinery and regulates both in unstressed conditions. These bivalent interactions allow KCTD10 to detect co-directional TRCs and facilitate higher-order assembly of KCTD10 complexes that can recruit CUL3 to ubiquitinate the RNA polymerase factor TCEA2. In the absence of KCTD10 there is increased retention of TCEA2 and RNA polymerase complex, causing an accumulation of TRCs and increased DNA damage. These data report results from a screen we performed using Alphafold2 to look for potential protein-protein interactions between KCTD10 and transcriptional regulators.