Zc3h12a is an RNase essential for controlling immune responses by regulating mRNA decay

Abstract

Toll-like receptors (TLRs) recognize microbial components, and evoke inflammation and immune responses1,2,3. TLR stimulation activates complex gene expression networks that regulate the magnitude and duration of the immune reaction. Here we identify the TLR-inducible gene Zc3h12a as an immune response modifier that has an essential role in preventing immune disorders. Zc3h12a-deficient mice suffered from severe anaemia, and most died within 12 weeks. Zc3h12a-/- mice also showed augmented serum immunoglobulin levels and autoantibody production, together with a greatly increased number of plasma cells, as well as infiltration of plasma cells to the lung. Most Zc3h12a-/- splenic T cells showed effector/memory characteristics and produced interferon-γ in response to T-cell receptor stimulation. Macrophages from Zc3h12a-/- mice showed highly increased production of interleukin (IL)-6 and IL-12p40 (also known as IL12b), but not TNF, in response to TLR ligands. Although the activation of TLR signalling pathways was normal, Il6 messenger RNA decay was severely impaired in Zc3h12a-/- macrophages. Overexpression of Zc3h12a accelerated Il6 mRNA degradation via its 3′-untranslated region (UTR), and destabilized RNAs with 3′-UTRs for genes including Il6, Il12p40 and the calcitonin receptor gene Calcr. Zc3h12a contains a putative amino-terminal nuclease domain, and the expressed protein had RNase activity, consistent with a role in the decay of Il6 mRNA. Together, these results indicate that Zc3h12a is an essential RNase that prevents immune disorders by directly controlling the stability of a set of inflammatory genes.

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Figure 1: Early onset of fetal autoimmune disease in Zc3h12a -/- mice.
Figure 2: Cellular abnormalities and augmented cytokine production in Zc3h12a -/- mice.
Figure 3: Zc3h12a destabilizes mRNA from a set of genes through their 3′-UTRs.
Figure 4: Zc3h12a contains RNase activity essential for destabilizing Il6 mRNA.

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Gene Expression Omnibus

Data deposits

Microarray data are deposited in the Gene Expression Omnibus (accession number GSE14890 for series of Myd88-/- and Trif-/- macrophages, and GSE14891 for series of Zc3h12a-/- macrophages). The structure model of Zc3h12a nuclease domain has been deposited in the Protein Model DataBase (PMDB) under accession number PM0075640.

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Acknowledgements

We thank all colleagues in our laboratory, E. Kamada for secretarial assistance, and Y. Fujiwara, M. Kumagai and R. Abe for technical assistance. We thank S. Sato for discussions and W. Zhao and K. Kirkwood for plasmids. This work was supported by the Special Coordination Funds of the Japanese Ministry of Education, Culture, Sports, Science and Technology, grants from the Ministry of Health, Labour and Welfare in Japan, the Global Center of Excellence Program of Japan, and the NIH (P01 AI070167).

Author Contributions K.M. generated Zc3h12a-/- mice and performed most experiments. O.T. identified Zc3h12a, designed the research and wrote the paper. D.M.S. and H.N. carried out structural modelling. Y.K. analysed microarray data, and T.T. was responsible for histological analysis. T.K., T.M., T.S. and H.K. helped with experiments. S.A. designed the research and supervised the project.

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Correspondence to Shizuo Akira.

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Matsushita, K., Takeuchi, O., Standley, D. et al. Zc3h12a is an RNase essential for controlling immune responses by regulating mRNA decay. Nature 458, 1185–1190 (2009). https://doi.org/10.1038/nature07924

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