Formation of co-transcriptional R-loops underlies replication fork stalling upon head-on transcription-replication encounters. Here, we demonstrate that RAD51-dependent replication fork reversal induced by R-loops is followed by the restart of semiconservative DNA replication mediated by RECQ1 and RECQ5 helicases, MUS81/EME1 endonuclease, RAD52 strand-annealing factor, the DNA ligase IV (LIG4)/XRCC4 complex, and the non-catalytic subunit of DNA polymerase δ, POLD3. RECQ5 disrupts RAD51 filaments assembled on stalled forks after RECQ1-mediated reverse branch migration, preventing a new round of fork reversal and facilitating fork cleavage by MUS81/EME1. MUS81-dependent DNA breaks accumulate in cells lacking RAD52 or LIG4 upon induction of R-loop formation, suggesting that RAD52 acts in concert with LIG4/XRCC4 to catalyze fork religation, thereby mediating replication restart. The resumption of DNA synthesis after R-loop-associated fork stalling also requires active transcription, the restoration of which depends on MUS81, RAD52, LIG4, and the transcription elongation factor ELL. These findings provide mechanistic insights into transcription-replication conflict resolution.
- Klíčová slova
- DNA ligase IV, MUS81, R-loop, RECQ5, replication fork reversal, replication restart, replication stress, transcription-replication conflict,
- MeSH
- DNA opravný a rekombinační protein Rad52 metabolismus MeSH
- DNA vazebné proteiny metabolismus MeSH
- DNA-ligasy metabolismus MeSH
- DNA-polymerasa III metabolismus MeSH
- endodeoxyribonukleasy metabolismus MeSH
- endonukleasy genetika metabolismus MeSH
- genetická transkripce genetika MeSH
- HeLa buňky MeSH
- helikasy RecQ metabolismus fyziologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- R-smyčka genetika fyziologie MeSH
- rekombinasa Rad51 genetika metabolismus fyziologie MeSH
- replikace DNA genetika fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA opravný a rekombinační protein Rad52 MeSH
- DNA vazebné proteiny MeSH
- DNA-ligasy MeSH
- DNA-polymerasa III MeSH
- endodeoxyribonukleasy MeSH
- endonukleasy MeSH
- helikasy RecQ MeSH
- Lig4 protein, Arabidopsis MeSH Prohlížeč
- MUS81 protein, human MeSH Prohlížeč
- RAD51 protein, human MeSH Prohlížeč
- RAD52 protein, human MeSH Prohlížeč
- RECQL protein, human MeSH Prohlížeč
- RECQL5 protein, human MeSH Prohlížeč
- rekombinasa Rad51 MeSH
- XRCC4 protein, human MeSH Prohlížeč
