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抗病毒蛋白 viperin 通过 CXCL10 蛋白分泌调节软骨分化。

The antiviral protein viperin regulates chondrogenic differentiation via CXCL10 protein secretion.

机构信息

From the Laboratory for Experimental Orthopedics, Department of Orthopedic Surgery and.

the Department of Anatomy and Embryology, Maastricht University, NL-6202 AZ Maastricht, The Netherlands.

出版信息

J Biol Chem. 2019 Mar 29;294(13):5121-5136. doi: 10.1074/jbc.RA119.007356. Epub 2019 Feb 4.

DOI:10.1074/jbc.RA119.007356
PMID:30718282
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6442052/
Abstract

Viperin (also known as radical SAM domain-containing 2 (RSAD2)) is an interferon-inducible and evolutionary conserved protein that participates in the cell's innate immune response against a number of viruses. mRNA is a substrate for endoribonucleolytic cleavage by RNase mitochondrial RNA processing (MRP) and mutations in the RNase MRP small nucleolar RNA (snoRNA) subunit of the RNase MRP complex cause cartilage-hair hypoplasia (CHH), a human developmental condition characterized by metaphyseal chondrodysplasia and severe dwarfism. It is unknown how CHH-pathogenic mutations in RNase MRP snoRNA interfere with skeletal development, and aberrant processing of RNase MRP substrate RNAs is thought to be involved. We hypothesized that viperin plays a role in chondrogenic differentiation. Using immunohistochemistry, real-time quantitative PCR, immunoblotting, ELISA, siRNA-mediated gene silencing, plasmid-mediated gene overexpression, label-free MS proteomics, and promoter reporter bioluminescence assays, we discovered here that viperin is expressed in differentiating chondrocytic cells and regulates their protein secretion and the outcome of chondrogenic differentiation by influencing transforming growth factor β (TGF-β)/SMAD family 2/3 (SMAD2/3) activity via C--C motif chemokine ligand 10 (CXCL10). Of note, we observed disturbances in this viperin-CXCL10-TGF-β/SMAD2/3 axis in CHH chondrocytic cells. Our results indicate that the antiviral protein viperin controls chondrogenic differentiation by influencing secretion of soluble proteins and identify a molecular route that may explain impaired chondrogenic differentiation of cells from individuals with CHH.

摘要

Viperin（也称为激进的 SAM 结构域包含 2（RSAD2））是一种干扰素诱导的进化保守蛋白，参与细胞对多种病毒的固有免疫反应。mRNA 是核糖核酸酶线粒体 RNA 加工（MRP）内切核糖核酸酶切割的底物，而核糖核酸酶 MRP 复合物中的核糖核酸酶 MRP 小核仁 RNA（snoRNA）亚基的突变会导致软骨毛发发育不良（CHH），这是一种人类发育状况，其特征为干骺端软骨发育不良和严重侏儒症。目前尚不清楚 CHH 相关的核糖核酸酶 MRP snoRNA 突变如何干扰骨骼发育，并且认为异常的核糖核酸酶 MRP 底物 RNA 加工与此有关。我们假设 viperin 在软骨生成分化中发挥作用。通过免疫组织化学，实时定量 PCR，免疫印迹，ELISA，siRNA 介导的基因沉默，质粒介导的基因过表达，无标记 MS 蛋白质组学和启动子报告基因生物发光测定，我们在这里发现 viperin 在分化的软骨细胞中表达，并通过影响转化生长因子β（TGF-β）/ SMAD 家族 2/3（SMAD2/3）活性来调节其蛋白质分泌和软骨生成分化的结果通过 C--C 基序趋化因子配体 10（CXCL10）。值得注意的是，我们观察到 CHH 软骨细胞中这种 viperin-CXCL10-TGF-β/SMAD2/3 轴的紊乱。我们的结果表明，抗病毒蛋白 viperin 通过影响可溶性蛋白的分泌来控制软骨生成分化，并确定了一条可能解释 CHH 患者细胞软骨生成分化受损的分子途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55a3/6442052/219c6a0c0d98/zbc0141903190009.jpg
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