SMARCAL1 ubiquitylation controls its association with RPA-coated ssDNA and promotes replication fork stability

The purpose of these experiments was to examine the genetic requirements for the replication stress induced by UV in RFWD3-depleted cells. We also examined the chromatin accumulation of SMARCAL1 in UV-treated RFWD3-depleted cells.
Conclusion:
RFWD3 depletion leads to high levels of RPA-ssDNA and y-H2.AX upon low-dose UV treatments and this is dependent on SMARCAL1, MUS81 but not on HLTF and ZRANB3. RFWD3 depletion also enhances chromatin recruitment of SMARCAL1 upon UV treatment. For each experiments non UV-treated samples were included to monitor baseline accumulation of RPA, SMARCAL1 and y-H2A.X on chromatin. In initial experiments, single stained controls were also included to confirm the absence of bleed-through.