TY - JOUR
T1 - The antiviral protein viperin regulates chondrogenic differentiation via CXCL10 protein secretion
AU - Steinbusch, Mandy M. F.
AU - Caron, Marjolein M. J.
AU - Surtel, Don A. M.
AU - van den Akker, Guus G. H.
AU - van Dijk, Paul J.
AU - Friedrich, Franziska
AU - Zabel, Bernhard
AU - van Rhijn, Lodewijk W.
AU - Peffers, Mandy J.
AU - Welting, Tim J. M.
N1 - Funding Information:
This work was supported by Bilateral Cooperation Grant DN 82-304 from The Netherlands Organisation for Scientific Research and the Deutsche Forsch-ungsgemeinschaft and by Dutch Arthritis Association Grant LLP14. This work is also supported in part by a Wellcome Trust Clinical Intermediate Fellowship Grant 107471 (to M. J. P.). The authors declare that they have no conflicts of interest with the contents of this article. We thank Prof. P. van der Kraan (Experimental Rheumatology, Radboud University Medical Center, Nijmegen, The Netherlands) for PAI1 and SMAD7 primer sequences; Prof. G. Pruijn (Biomolecular Chemistry, Radboud University, Nijmegen, The Netherlands) for discussion; Dr. T. Dalmay (School of Biological Sciences, University of East Anglia, Norwich, United Kingdom) for the CAGA12-luciferase reporter plasmid; Prof. P. Cresswell (Department of Immunobiology, Yale University, New Haven, CT) for a plasmid containing the viperin ORF; Dr. A. Groot (Maastro, Maastricht University, Maastricht, The Netherlands) for the pGluc-CMV plasmid; and Dr. S. Köhler (Anatomy and Embryology, Maastricht University, Maastricht, The Netherlands) for assistance with paraffin sectioning.
Publisher Copyright:
© 2019 Steinbusch et al. Published by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2019/3/29
Y1 - 2019/3/29
N2 - 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. Viperin 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 beta (TGF-beta)/SMAD family 2/3 (SMAD2/3) activity via C-X-C motif chemokine ligand 10 (CXCL10). Of note, we observed disturbances in this viperin-CXCL10-TGF-beta/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.
AB - 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. Viperin 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 beta (TGF-beta)/SMAD family 2/3 (SMAD2/3) activity via C-X-C motif chemokine ligand 10 (CXCL10). Of note, we observed disturbances in this viperin-CXCL10-TGF-beta/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.
KW - cartilage biology
KW - chondrogenesis
KW - small nucleolar RNA (snoRNA)
KW - transforming growth factor beta (TGF-B)
KW - chemokine
KW - secretion
KW - C-X-C motif chemokine ligand 10
KW - cartilage-hair hypoplasia
KW - chondrogenic differentiation
KW - RMRP snoRNA
KW - viperin
KW - INTERFERON-INDUCIBLE PROTEIN
KW - MESENCHYMAL STEM-CELLS
KW - RNASE MRP
KW - GENE-EXPRESSION
KW - MESSENGER-RNA
KW - GROWTH-FACTOR
KW - INHIBITION
KW - GAMMA
KW - ACTIVATION
KW - MUTATIONS
U2 - 10.1074/jbc.RA119.007356
DO - 10.1074/jbc.RA119.007356
M3 - Article
C2 - 30718282
SN - 0021-9258
VL - 294
SP - 5121
EP - 5136
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 13
ER -