RRC ID 67087
Author Jokoji G, Maeda S, Oishi K, Ijuin T, Nakajima M, Tawaratsumida H, Kawamura I, Tominaga H, Taketomi E, Ikegawa S, Taniguchi N.
Title CDC5L promotes early chondrocyte differentiation and proliferation by modulating pre-mRNA splicing of SOX9, COL2A1, and WEE1.
Journal J Biol Chem
Abstract Ossification of the posterior longitudinal ligament (OPLL) of the spine is a common pathological condition that causes intractable myelopathy and radiculopathy, mainly the result of an endochondral ossification-like process. Our previous genome-wide association study identified six susceptibility loci for OPLL, including the CDC5L gene region. Here, we found CDC5L to be expressed in type II collagen-producing chondrocyte-like fibroblasts in human OPLL specimens, as well as in differentiating ATDC5 chondrocytes. Cdc5l siRNA transfection in murine chondrocytes decreased the expression of the early chondrogenic genes Sox9 and Col2a1, diminished the cartilage matrix production, and enhanced the expression of parathyroid hormone-related protein (a resting chondrocyte marker). We also showed that Cdc5l shRNA suppressed the growth of cultured murine embryonal metatarsal cartilage rudiments, and that Cdc5l knockdown suppressed the growth of ATDC5 cells. Fluorescence-activated cell sorting analysis revealed that the G2/M cell cycle transition was blocked; our data showed that Cdc5l siRNA transfection enhanced expression of Wee1, an inhibitor of the G2/M transition. Cdc5l siRNA also decreased the pre-mRNA splicing efficiency of Sox9 and Col2a1 genes in both ATDC5 cells and primary chondrocytes; conversely, loss of Cdc5l resulted in enhanced splicing of Wee1 pre-mRNA. Finally, an RNA-binding protein immunoprecipitation assay revealed that Cdc5l bound directly to these target gene transcripts. Overall, we conclude that Cdc5l promotes both early chondrogenesis and cartilage growth, and may play a role in the etiology of OPLL, at least in part by fine-tuning the pre-mRNA splicing of chondrogenic genes and Wee1, thus initiating the endochondral ossification process.
Volume 297(2)
Pages 100994
Published 2021-8-1
DOI 10.1016/j.jbc.2021.100994
PII S0021-9258(21)00796-1
PMID 34298017
PMC PMC8363834
MeSH Animals Cell Cycle Proteins / genetics Cell Cycle Proteins / metabolism* Cell Differentiation Cell Line Chondrocytes / cytology* Chondrocytes / metabolism Chondrogenesis* Collagen Type II / genetics Collagen Type II / metabolism* Humans Mice Models, Animal Osteogenesis / physiology Protein-Tyrosine Kinases / genetics Protein-Tyrosine Kinases / metabolism* RNA Splicing* RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism* SOX9 Transcription Factor / genetics SOX9 Transcription Factor / metabolism*
IF 4.238
Human and Animal Cells ATDC5(RCB0565)