JMJD3 regulates CD4+ T cell trafficking by targeting actin cytoskeleton regulatory gene Pdlim4
Chuntang Fu, … , Helen Y. Wang, Rong-Fu Wang
Chuntang Fu, … , Helen Y. Wang, Rong-Fu Wang
Published August 8, 2019
Citation Information: J Clin Invest. 2019;129(11):4745-4757. https://doi.org/10.1172/JCI128293.
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Research Article Autoimmunity Cell biology Immunology

JMJD3 regulates CD4+ T cell trafficking by targeting actin cytoskeleton regulatory gene Pdlim4

Abstract

Histone H3K27 demethylase JMJD3 plays a critical role in gene expression and T cell differentiation. However, the role and mechanisms of JMJD3 in T cell trafficking remain poorly understood. Here, we show that JMJD3 deficiency in CD4+ T cells resulted in an accumulation of T cells in the thymus and reduction of T cell number in the secondary lymphoid organs. We identified PDLIM4 as a significantly downregulated target gene in JMJD3-deficient CD4+ T cells by gene profiling and ChIP-Seq analyses. We further showed that PDLIM4 functioned as an adaptor protein to interact with sphingosine-1 phosphate receptor 1 (S1P1) and filamentous actin (F-actin), thus serving as a key regulator of T cell trafficking. Mechanistically, JMJD3 bound to the promoter and gene-body regions of the Pdlim4 gene and regulated its expression by interacting with zinc finger transcription factor KLF2. Our findings have identified Pdlim4 as a JMJD3 target gene that affects T cell trafficking by cooperating with S1P1 and have provided insights into the molecular mechanisms by which JMJD3 regulates genes involved in T cell trafficking.

Authors

Chuntang Fu, Qingtian Li, Jia Zou, Changsheng Xing, Mei Luo, Bingnan Yin, Junjun Chu, Jiaming Yu, Xin Liu, Helen Y. Wang, Rong-Fu Wang

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Figure 5

JMJD3 regulates Pdlim4 expression by interacting with KLF2.

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JMJD3 regulates Pdlim4 expression by interacting with KLF2. (A) Screenin...
(A) Screening of transcription factors interacting with JMJD3. 293T cells were cotransfected with HA-tagged Runx2, Klf4, Klf2, Wdr5, Fosl1, FosB, Wnt5a, Nkx2.1, and FLAG-Jmjd3. WCLs were immunoprecipitated with anti-FLAG Abs and immunoblotted with anti-HA Abs. (B) Schematic presentation of the Pdlim4 promoter–driven luciferase construct. The promoter region –1512 bp upstream was cloned into an episomal luciferase vector. (C) The transcriptional activity of proteins interacting with JMJD3 in regulating Pdlim4 was evaluated by dual-luciferase assay. Data are presented as mean ± SD from 3 independent experiments (n = 3). **P < 0.01, 1-way ANOVA with Tukey’s multiple comparisons test. (D) Thymic CD4 SP T cells were isolated from WT and Jmjd3-cKO mice and immunoprecipitated with anti-JMJD3 or anti-KLF2 Abs and protein (A+G) beads. The immunoprecipitated product was immunoblotted with anti-KLF2 or anti-JMJD3 Abs. (E) Mapping the KLF2 and JMJD3 binding regions of the Pdlim4 promoter using a dual-luciferase assay. Different regions of the Pdlim4 promoter were cloned into the episomal luciferase vector and then were cotransfected with Klf2 and Jmjd3 into 293T cells. Data are presented as mean ± SD from 3 independent experiments (n = 3). *P < 0.05, Student’s t test. (F) ChIP-qPCR analysis of percentage of enrichment of KLF2 at the Pdlim4 promoter in WT and Jmjd3-cKO thymic CD4 SP T cells after ectopic expression. Jmjd3 IgG was used as isotype control. n = 3. **P < 0.01, 1-way ANOVA with Tukey’s multiple comparisons test.