A role for Rad5 in ribonucleoside monophosphate (rNMP) tolerance

Abstract

Ribonucleoside monophosphate (rNMP) incorporation in genomic DNA poses a significant threat to genomic integrity. In addition to repair, DNA damage tolerance mechanisms ensure replication progression upon encountering unrepaired lesions. One player in the tolerance mechanism is Rad5, which is an E3 ubiquitin ligase and helicase. Here, we report a new role for yeast Rad5 in tolerating rNMP incorporation, in the absence of the bona fide ribonucleotide excision repair pathway via RNase H2. This role of Rad5 is further highlighted after replication stress induced by hydroxyurea or by increasing rNMP genomic burden using a mutant DNA polymerase (Pol ε - Pol2-M644G). We further demonstrate the importance of the ATPase and ubiquitin ligase domains of Rad5 in rNMP tolerance. These findings suggest a similar role for the human Rad5 homologues helicase-like transcription factor (HLTF) and SNF2 Histone Linker PHD RING Helicase (SHPRH) in rNMP tolerance, which may impact the response of cancer cells to replication stress-inducing therapeutics.

Genetic requirements for repair of lesions caused by single genomic ribonucleotides in S phase

“…2B ), we switched to HU instead of pol2-M644G to modulate the genomic rNMP load. HU has been previously shown to increase the genomic rNMP load in yeast and mammalian cells 23 , 24 . As expected, in the presence of HU, the S-RNH202 allele was synthetic sick with rtt101 Δ (Supplementary Fig.…”