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.
View: Text | PDF
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

×

Figure 1

The human MIF gene and the strategy for the identification of MIF –794 CATT5–8–interacting proteins.

Options: View larger image (or click on image) Download as PowerPoint
The human MIF gene and the strategy for the identification of MIF –794 C...
(A) Diagram illustrating the 3 MIF exons, predicted transcription factor–binding sites, and the –794 CATT5–8 microsatellite repeat (rs5844572). The numerical prefixes refer to nucleotide distance (in bp) upstream from the transcription start site. (B) Diagram of synthetic 5′ biotin-labeled oligonucleotides spanning nucleotides –865/–833 to –752 of the MIF promoter used for differential affinity chromatography of human THP-1 monocyte nuclear proteins. The corresponding double-stranded oligos were used experimentally. (C) Verification of retention of the nuclear transcription factor Pit-1 by the 5′CATT8 but not 5′CATT0 oligonucleotide. Recombinant Pit-1 (10 ng) incubated with 100 nM MIF promoter 5′CATT0 or 5′CATT8 oligos prior to the addition of streptavidin beads, NaCl elution, and SDS-PAGE (4%–12%) of eluates, followed by Western blotting with anti–Pit-1 (lanes 1 and 2). Positive control Western blot showing recombinant Pit-1 alone (no oligonucleotide addition) (lane 3). The blot is representative of 3 experiments.