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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Research Article Immunology

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 4

Regulation of TLR-activated MIF expression in human monocytes by ICBP90.

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Regulation of TLR-activated MIF expression in human monocytes by ICBP90....
THP-1 monocytes were transfected with MIF promoter-luciferase reporter plasmids bearing 0, 5, 6, 7, and 8 CATT repeats and treated with an ICBP90 or control shRNA, cultured for 6 hours, and (A) were left unstimulated or were stimulated with (B) Pam3CysK (100 ng/ml) for TLR1/2 agonism, (C) polyI:C (1 μg/ml) for TLR3 agonism, (D) LPS (100 ng/ml) for TLR4 agonism, (E) flagellin (100 ng/ml) for TLR5 agonism, or (F) CpG DNA (5 μM) for TLR9 agonism prior to measurement of luciferase activity. Primary human peripheral blood monocytes were similarly analyzed under (G) basal and (H) LPS-stimulated conditions (100 ng/ml). 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 vs. ICBP90 shRNA within each panel; #P < 0.05, ##P < 0.01 for stimulated vs. basal expression for B–F vs. A and for H vs. G (2-tailed Student’s t test).