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 4

PDLIM4 regulates T cell migration by interaction with S1P1 and modulation of F-actin reorganization.

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PDLIM4 regulates T cell migration by interaction with S1P1 and modulatio...
(A) Immunofluorescence microscopy images of WT and Jmjd3-cKO CD4+ T cells infected with lentivirus containing GFP or PDLIM4-GFP plasmids. Cells were starved 12 hours and treated with 100 nM S1P for 3 hours at 37°C. GFP or PDLIM4-GFP was detected with green fluorescence. Actin filaments were labeled with rhodamine-conjugated phalloidin (red). Nuclei were stained with DAPI (blue). (B) FACS analysis of phalloidin-labeled F-actin in untreated and S1P-treated WT, Jmjd3-cKO, Pdlim4-KO, and Pdlim4-expressing Jmjd3-cKO CD4 SP thymocytes. (C) F-actin (pellet [P]) and G-actin (supernatant [S]) from untreated and treated WT, Jmjd3-cKO CD4 SP thymocytes were detected by Western blot. (D) Immunofluorescence microscopy of untreated and S1P-treated WT CD4+ SP cells stained with FITC-conjugated Abs to detect PDLIM4, Cy5-conjugated Abs to detect S1P1, and rhodamine-conjugated phalloidin to detect actin filaments. Goat IgG and rabbit IgG were used as isotype controls. Merged images indicate colocalization of proteins. (E) Co-IP analysis of endogenous interaction of PDLIM4 with S1P1 in untreated and S1P-treated CD4 SP thymocytes. (F) Cosedimentation assay was performed using GST-PDLIM4, GST-PDLIM4 N-del, or GST-PDLIM4 C-del with F-actin, and subsequent analysis of supernatants and pellets by Western blot analysis. (G) 293T cells were cotransfected with FLAG-Pdlim4-N-del, C-del, or full-length Pdlim4 along with HA-S1p1. WCLs were immunoprecipitated with anti-HA beads and immunoblotted with anti-FLAG Ab. Three independent experiments were repeated with similar results. (H) A schematic diagram of the proposed model showing that PDLIM4 interacts with the S1P1 protein at the N-terminal PDZ domain and binds F-actin by the C-terminal LIM domain.