Language
English
Publication Date
10-11-2024
Journal
bioRxiv
DOI
10.1101/2024.10.10.617668
PMID
39416207
PMCID
PMC11482920
PubMedCentral® Posted Date
10-11-2024
PubMedCentral® Full Text Version
Pre-Print
Abstract
Diamond Blackfan anemia (DBA) is caused by germline heterozygous loss-of-function pathogenic variants (PVs) in ribosomal protein (RP) genes, most commonly RPS19 and RPL5. In addition to red cell aplasia, individuals with DBA are at increased risk of various cancers. Importantly, the mechanism(s) underlying cancer predisposition are poorly understood. We found that DBA patient-derived lymphoblastoid cells had persistent γ-H2AX foci following ionizing radiation (IR) treatment, suggesting DNA double-strand break (DSB) repair defects. RPS19- and RPL5-knocked down (KD) CD34+ cells had delayed repair of IR-induced DSBs, further implicating these RPs in DSB repair. Assessing the impact of RPS19- and RPL5-KD on specific DSB repair pathways, we found RPS19-KD decreased the efficiency of pathways requiring extensive end-resection, whereas RPL5-KD increased end-joining pathways. Additionally, RAD51 was reduced in RPS19- and RPL5-KD and RPS19- and RPL5-mutated DBA cells, whereas RPS19-deficient cells also had a reduction in PARP1 and BRCA2 proteins. RPS19-KD cells had an increase in nuclear RPA2 and a decrease in nuclear RAD51 foci post-IR, reflective of alterations in early, critical steps of homologous recombination. Notably, RPS19 and RPL5 interacted with poly(ADP)-ribose chains noncovalently, were recruited to DSBs in a poly(ADP)-ribose polymerase activity-dependent manner, and interacted with Ku70 and histone H2A. RPL5’s recruitment, but not RPS19’s, also required p53, suggesting that RPS19 and RPL5 directly participate in DSB repair via different pathways. We propose that defective DSB repair arising from haploinsufficiency of these RPs may underline the cancer predisposition in DBA.
Published Open-Access
yes
Recommended Citation
DeCleene, Nicholas F; Asik, Elif; Sanchez, Anthony; et al., "RPS19 and RPL5, the Most Commonly Mutated Genes in Diamond Blackfan Anemia, Impact Dna Double-Strand Break Repair" (2024). Faculty, Staff and Students Publications. 8536.
https://digitalcommons.library.tmc.edu/baylor_docs/8536