Transcription factor ICBP90 regulates the MIF promoter and immune susceptibility locus
Jie Yao, … , Patty Lee, Richard Bucala
Jie Yao, … , Patty Lee, Richard Bucala
Published January 11, 2016
Citation Information: J Clin Invest. 2016;126(2):732-744. https://doi.org/10.1172/JCI81937.
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Transcription factor ICBP90 regulates the MIF promoter and immune susceptibility locus

Abstract

The immunoregulatory cytokine macrophage migration inhibitory factor (MIF) is encoded in a functionally polymorphic locus that is linked to the susceptibility of autoimmune and infectious diseases. The MIF promoter contains a 4-nucleotide microsatellite polymorphism (–794 CATT) that repeats 5 to 8 times in the locus, with greater numbers of repeats associated with higher mRNA levels. Because there is no information about the transcriptional regulation of these common alleles, we used oligonucleotide affinity chromatography and liquid chromatography–mass spectrometry to identify nuclear proteins that interact with the –794 CATT5–8 site. An analysis of monocyte nuclear lysates revealed that the transcription factor ICBP90 (also known as UHRF1) is the major protein interacting with the MIF microsatellite. We found that ICBP90 is essential for MIF transcription from monocytes/macrophages, B and T lymphocytes, and synovial fibroblasts, and TLR-induced MIF transcription is regulated in an ICBP90- and –794 CATT5–8 length–dependent manner. Whole-genome transcription analysis of ICBP90 shRNA–treated rheumatoid synoviocytes uncovered a subset of proinflammatory and immune response genes that overlapped with those regulated by MIF shRNA. In addition, the expression levels of ICBP90 and MIF were correlated in joint synovia from patients with rheumatoid arthritis. These findings identify ICBP90 as a key regulator of MIF transcription and provide functional insight into the regulation of the polymorphic MIF locus.

Authors

Jie Yao, Lin Leng, Maor Sauler, Weiling Fu, Junsong Zheng, Yi Zhang, Xin Du, Xiaoqing Yu, Patty Lee, Richard Bucala

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Figure 5

Comparative effect of ICBP90 versus MIF knockdown on MIF-dependent cytokine production.

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Comparative effect of ICBP90 versus MIF knockdown on MIF-dependent cytok...
Human THP-1 monocytes were treated with shRNA directed against ICBP90, MIF, or an shRNA control and stimulated for 6 hours with LPS (100 ng/ml). Conditioned media then were harvested for analysis by specific ELISA for (A) MIF, (B) TNF-α, (C) IL-1β, (D) IL-6, (E) IL-8, and (F) MCP-1. Data are presented as the mean+SD of 3 measurements, with all experiments replicated twice (n = 3 measurements per experiment). *P < 0.05, **P < 0.01 for control shRNA versus MIF shRNA as well as for control shRNA versus ICBP90 shRNA by 1-way ANOVA for repeated measurements followed by Dunnett’s test for comparing the 2 knockdown groups with the control group at individual time points. P values are shown only for those comparisons that were significantly different.