A Novel FANCD2-Chromatin Bound Complex in Replication Stress Response and HSC Maintenance
Abstract
Due to the ongoing COVID-19 pandemic, we faced repeated disruption of our research activities in the past several months. However, we have mostly finished Aims 1 and 2 in the second funding year. We have generated the Fanca-KO;Fancd2-KI mice, which are deficient for the essential component of the FA core complex (Fanca). We have performed preliminary experiments to determine whether expression of the 3XFLAG-Fancd2 KI allele in Fanca-KO mice would alter the FA hallmark phenotype; DNA damage and homologous recombination (HR) repair. We isolated LSK (Lin-Sca1+c-kit+; enriched for HSCs) from Fanca-KO;Fancd2-KI mice and the WT;Fancd2-KI control mice, and exposed the cells to MMC (5 ng/ml) to assess MMC-induced H2AX and RAD51 foci formation. We observed a significant increase in MMC-induced formation of -H2AX foci in Fanca-KO;Fancd2- KI LSK cells compared to those from the WT;Fancd2-KI control. MMC-induced HR repair, as determined by the formation of RAD51 foci, was evidently defective in Fanca-KO;Fancd2-KI LSK cells. Our studies on the functional consequence of the interaction between the FANCD2-RNF40 and FANCD2-EDD has the potential to establish the functional link between the FANCD2/FA pathway and replication stress response in genomic maintenance. We also expect these functional data to uncover whether these interacting pathways may be viable targets for therapy in FA BM failure.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2022
- Accession Number
- AD1215423
Entities
People
- Ruhikanta Meetei