Project description:Premature ovarian insufficiency (POI) is a heterogeneous female disorder characterized by the loss of ovarian function before the age of 40. It represents a significant detriment to female fertility. However, the known POI-causative genes currently account for only a fraction of cases. To elucidate the genetic factors underlying POI, we conducted whole-exome sequencing on a family with three fertile POI patients and identified a deleterious missense variant in RNF111. In a subsequent replication study involving 1030 POI patients, this variant was not only confirmed but also accompanied by the discovery of three additional predicted deleterious RNF111 variants. These variants collectively account for eight cases, representing 0.78% of the study cohort. A further study involving 500 patients with diminished ovarian reserves also identified two additional RNF111 variants. Notably, RNF111 encodes an E3-ubiquitin ligase with a regulatory role in the TGF-β/BMP signaling pathway. Our analysis revealed that RNF111/RNF111 is predominantly expressed in the oocytes of mice, monkeys, and humans. To further investigate the functional implications of RNF111 variants, we generated two mouse models: one with a heterozygous missense mutation (Rnf111+/M) and another with a heterozygous null mutation (Rnf111+/−). Both mouse models exhibited impaired female fertility, characterized by reduced litter sizes and small ovarian reserve. Additionally, RNA-seq and quantitative proteomics analysis unveiled that Rnf111 haploinsufficiency led to dysregulation in female gonad development and negative regulation of the BMP signaling pathway within mouse ovaries. In conclusion, our findings strongly suggest that monoallelic deleterious variants in RNF111 can impair female fertility and induce POI in both humans and mice.
Project description:Premature ovarian insufficiency (POI) is a heterogeneous female disorder characterized by the loss of ovarian function before the age of 40. It represents a significant detriment to female fertility. However, the known POI-causative genes currently account for only a fraction of cases. To elucidate the genetic factors underlying POI, we conducted whole-exome sequencing on a family with three POI patients and identified a deleterious missense variant in RNF111. In a subsequent replication study involving 1030 POI patients, this variant was not only confirmed but also accompanied by the discovery of three additional predicted deleterious RNF111 variants. These variants collectively account for eight cases, representing 0.78% of the study cohort. A further study involving 500 patients with diminished ovarian reserves also identified two additional RNF111 variants. Notably, RNF111 encodes an E3-ubiquitin ligase with a regulatory role in the TGF-β/BMP signaling pathway. Our analysis revealed that RNF111/RNF111 is
Project description:To investigate the potential impact of VHL on chromatin accessibility, the assay for transposase-accessible chromatin with high throughput sequencing (ATAC-seq) was performed in renal cell carcinoma cells.The VHL wild-type Caki-1 cells were used to generate control or VHL knockout cells by CRISPR/Cas9-mediated genome editing (sgCTR and sgVHL Caki-1 cells).Following ATAC-seq, DNAs were amplified using non-biased conditions, labeled, and sequenced with Illumina NovaSeq 6000.
Project description:In this study we performed 2 interactomes. To identify interactants of RNF111 that are dependent of the RING domain, we performed qualitative interactome comparison of HEK-293 cells transfected with GFP, GFP-RNF111-wt or GFP-RNF11-C933A mutated in its RING domain. This led to the identification of UBXN7 as a RING-dependent partner for RNF111. To identify UBXN7 UAS dependant partners, we performed quantitative interactome comparison of HEK-293 cells transfected with GFP, GFP-UBXN7-UAS.
Project description:Dissection of the functional input of transcription factors in embryos at a molecular and cellular level by sci-ATAC-seq profiling wild-type and TF mutant embryos on a time course of mesoderm/muscle development.
