Fork Cleavage-Religation Cycle and Active Transcription Mediate Replication Restart after Fork Stalling at Co-transcriptional R-Loops

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 R...

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Published in:Molecular cell Vol. 77; no. 3; p. 528
Main Authors: Chappidi, Nagaraja, Nascakova, Zuzana, Boleslavska, Barbora, Zellweger, Ralph, Isik, Esin, Andrs, Martin, Menon, Shruti, Dobrovolna, Jana, Balbo Pogliano, Chiara, Matos, Joao, Porro, Antonio, Lopes, Massimo, Janscak, Pavel
Format: Journal Article
Language:English
Published: United States 06.02.2020
Subjects:
Cell Line, Tumor
DNA Ligases - metabolism
DNA Polymerase III - metabolism
DNA Replication - genetics
DNA Replication - physiology
DNA-Binding Proteins - metabolism
Endodeoxyribonucleases - metabolism
Endonucleases - genetics
Endonucleases - metabolism
HeLa Cells
Humans
R-Loop Structures - genetics
R-Loop Structures - physiology
Rad51 Recombinase - genetics
Rad51 Recombinase - metabolism
Rad51 Recombinase - physiology
Rad52 DNA Repair and Recombination Protein - metabolism
RecQ Helicases - metabolism
RecQ Helicases - physiology
Transcription, Genetic - genetics
MUS81
replication stress
transcription-replication conflict
R-loop
replication restart
RECQ5
DNA ligase IV
replication fork reversal
ISSN:1097-4164, 1097-4164
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Summary: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.
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ISSN:1097-4164
1097-4164
DOI:10.1016/j.molcel.2019.10.026