bioRxiv Science⌕ Search

Biology subjects

Bishara, L. A.

Publications and source records attributed to Bishara, L. A..

2 recordsLinked to original sources

LSm4 biomolecular condensates drive XRN2-mediated RNA decay at DNA double-strand breaks to facilitate repair

Maintenance of genome integrity requires accurate repair of DNA double-strand breaks (DSBs), particularly within transcriptionally active regions. Persistent R-loops at DSBs can impede homologous recombination (HR) repair. While factors that resolve R-loops at DSB sites are known, the mechanisms ensuring timely degradation of nascent RNA to prevent pathological R-loop accumulation remain elusive. Here, we identified a critical role for the RNA-binding protein LSm4 in orchestrating localized RNA decay at DSBs to facilitate repair. We demonstrated that among LSm1-8 subunits, only LSm4 undergoes liquid-liquid phase separation (LLPS) and forms biomolecular condensates (BCs) specifically at DSBs in transcriptionally active chromatin. These damage-induced LSm4 BCs function as hubs that promote nuclear RNA decapping and recruit the 5'[->]3' exonuclease XRN2 to degrade nascent transcripts proximal to DSBs. Accordingly, LSm4-XRN2 axis suppresses R-loop hyperaccumulation, thereby enabling efficient RAD51 filament assembly and intact HR repair. Consequently, loss of LSm4 increases translocations and leads to genomic instability. Collectively, our findings define a new regulatory layer in which LSm4 BCs spatially license RNA degradation, preventing R-loop accumulation at DSB microenvironment to facilitate error-free repair.

molecular biology↗

Systematic identification of a CDYL1-dependent decrease in lysine crotonylation at DNA double-strand break sites functionally uncouples transcriptional silencing and repair

Previously, we showed that CDYL1 is recruited to DNA double-strand breaks (DSBs) to promote homology-directed repair (HDR) and foster transcriptional silencing. Yet, how CDYL1 elicits DSB-induced silencing is not fully understood. Here, we systematically identify a CDYL1-dependent local decrease in the transcriptionally active marks lysine crotonylation (PanKcr) and crotonylated histone residue H3K9cr at AsiSI-induced DSBs, which correlates with transcriptional silencing. Mechanistically, we reveal that CDYL1 crotonyl-CoA hydratase activity counteracts PanKcr and H3K9cr at AsiSI sites, which triggers the eviction of the transcriptional elongation factor ENL and foster transcriptional silencing. Furthermore, genetic inhibition of CDYL1 hydratase activity blocks the reduction in H3K9cr and alleviates DSB-induced silencing, while HDR efficiency unexpectedly remains intact. Therefore, our results functionally uncouple the repair and silencing activity of CDYL1 at DSBs. In a broader context, we address a long-standing question concerning the functional relationship between HDR and DSB-induced transcriptional silencing, suggesting that they may occur independently. HighlightsO_LISystematic identification of a local decrease in lysine crotonylation PanKcr and H3K9cr at AsiSI-induced DSBs that correlates with transcriptional silencing. C_LIO_LICDYL1 crotonyl-CoA hydratase activity downregulates Kcr at DSBs. C_LIO_LIKcr reduction at DSBs promotes ENL eviction and fosters transcriptional silencing. C_LIO_LICDYL1 roles in DSB-induced transcriptional silencing and HDR are functionally uncoupled. C_LI

molecular biology↗