Uhrf1 is indispensable for normal limb growth by regulating chondrocyte differentiation through specific gene expression

Abstract

Transcriptional regulation can be tightly orchestrated by epigenetic regulators. Among these, ubiquitin-like with PHD and RING finger domains 1 (Uhrf1) is reported to have diverse epigenetic functions, including regulation of DNA methylation. However, the physiological functions of Uhrf1 in skeletal tissues remain unclear. Here, we show that limb mesenchymal cell-specific Uhrf1 conditional knockout mice ( ) exhibit remarkably shortened long bones that have morphological deformities due to dysregulated chondrocyte differentiation and proliferation. RNA-seq performed on primary cultured chondrocytes obtained from mice showed abnormal chondrocyte differentiation. In addition, integrative analyses using RNA-seq and MBD-seq revealed that Uhrf1 deficiency decreased genome-wide DNA methylation and increased gene expression through reduced DNA methylation in the promoter regions of 28 genes, including, which is reported to be an IL1-related gene and to affect chondrocyte differentiation. knockdown in cKO chondrocytes can normalize abnormal expression of genes involved in chondrocyte differentiation, such as These results indicate that Uhrf1 governs cell type-specific transcriptional regulation by controlling the genome-wide DNA methylation status and regulating consequent cell differentiation and skeletal maturation.

GPRC5A facilitates cell proliferation through cell cycle regulation and correlates with bone metastasis in prostate cancer

“…Tibial bone mineral density (BMD) was assessed by dual‐energy X‐ray absorption measurements using a bone mineral analyzer (DCS‐600 EX Aloka, Japan). Micro computerized tomography scan was performed using μCT35 system (SCANCO Medical, Switzerland) as previously described …”

“…Micro computerized tomography scan was performed using μCT35 system (SCANCO Medical, Switzerland) as previously described. 28…”

“…Total RNA was collected from Control PC3-Luc and the two GPRC5A KO cell lines as described above and RNA-seq was performed as previously described. 28 Mapping to human genome data hg38 was performed using Tophat, and the expression analysis tool Cufflinks was used to normalize each sample before determining the differences in expression between Control (n = 3) and GPRC5A KO 1 and KO 2 (n = 6) by tmm using R. Raw and processed data were registered to the GEO (GSE121319). Gene Ontology analyses were performed using Database for Annotation, Visualization and Integrated Discovery (DAVID) Bioinformatics Resources 6.7 30 and Gene Set Enrichment Analysis (GSEA).…”

Earlier demethylation of myogenic genes contributes to embryonic precocious terminal differentiation of myoblasts in miniature pigs

“…Several methods, including bisulfite sequencing (BSseq), reduced representation BS-seq (RRBS) (17,18), methylated DNA immunoprecipitation (MeDIP-seq) (19), methyl-binding domain sequencing (20,21), and Bead-Chip array (22) have been developed to analyze patterns of DNA methylation across diverse vertebrate genomes, such as mouse whole-genome BS-seq (WGBS) (23), pig RRBS (24), bovine MeDIP-seq (25), zebrafish methylbinding domain sequencing (26) and chicken BeadChip array (27), etc. However, RRBS reduces the portion of the genome analyzed through MspI digestion and fragment size selection, but it is less efficient when analyzing tissue samples and requires much deeper sequence coverage (28).…”

UHRF1-repressed 5’-hydroxymethylcytosine is essential for the male meiotic prophase I

“…They also pointed out that the DNA methylation might regulate directly or indirectly the genes guiding spermatogonial differentiation (for example, Plzf and Kit) 23 . One of the key player of DNA methylation is UHRF1, a multifunctional protein [24][25][26] , which is essential for maintenance or de novo DNA methylation [27][28][29][30][31][32][33][34] . However, the role of UHRF1 in male germ cells after the spermatogonia differentiation, for example, meiosis, remains largely unknown.…”