Elsevier

Cellular Signalling

Volume 67, March 2020, 109522
Cellular Signalling

TRIM59 expression is regulated by Sp1 and Nrf1 in LPS-activated macrophages through JNK signaling pathway

https://doi.org/10.1016/j.cellsig.2019.109522Get rights and content

Highlights

  • TRIM59 is highly expressed in macrophages;

  • TRIM59 is markedly decreased by LPS stimuli in vivo and in vitro;

  • TRIM59 expression is regulated by Sp1 and Nrf1 in LPS-activated macrophages through JNK signaling pathway;

  • TRIM59 expression is decreased during atherosclerosis progression.

Abstract

Activated macrophages play an important role in many inflammatory diseases including septic shock and atherosclerosis. TRIM59 has been showed to participate in many pathological processes, such as inflammation, cytotoxicity and tumorigenesis. However, the molecular mechanisms controlling its expression in activated macrophages are not fully understood. Here we report that TRIM59 expression is regulated by Sp1 and Nrf1 in LPS-activated macrophages. TRIM59 is highly expressed in macrophages, and markedly decreased by LPS stimuli in vivo and in vitro. TRIM59 promoter activity is also significantly suppressed by LPS and further analysis demonstrated that Sp1 and Nrf1 directly bound to the proximal promoter of TRIM59 gene. LPS treatment significantly decreased Sp1 expression, nuclear translocation and reduced its binding to the promoter, whereas increased Nrf1 expression, nuclear translocation and enhanced its binding to the promoter. Moreover, LPS-decreased TRIM59 expression was reversed by JNK inhibitor. Finally, TRIM59 level is significantly decreased during atherosclerosis progression. Taken together, our results demonstrated that TRIM59 expression was precisely regulated by Sp1 and Nrf1 in LPS-activated macrophages, which may be dependent on the activation of JNK signaling pathway and TRIM59 may be a potential therapeutic target for inflammatory diseases such as atherosclerosis.

Introduction

Inflammation is the initial host immune reaction and is mediated by activating various immune cells, such as macrophages, which play an important role in host defense against microbial and viral infections [1,2]. Activated macrophages by inflammatory stimuli elaborate a large array of cytokines, growth factors and proteolytic enzymes that are critical for tissue damage and repair [[3], [4], [5]]. However, overproduction of these inflammatory mediators may lead to pathogenesis of many inflammatory diseases, such as septic shock, cancer and atherosclerosis [6,7]. Atherosclerosis is recognized as a chronic inflammatory disease and is the primary cause of most cardiovascular diseases, such as coronary artery disease (CAD), myocardial infarctions and strokes [8,9]. As the major component in atherosclerotic lesions, macrophages play a critical role in the development of atherosclerosis [10].

Lipopolysaccharide (LPS/endotoxin), the major constituent of the outer membrane of gram-negative bacteria, was used to induce activation of macrophages experimentally [2]. Mechanically, LPS mainly interacts with monocytes and/or macrophages via Toll-like receptor 4 (TLR4) to activate a number of intracellular signal pathways including PI3K/Akt, mitogen-activated protein kinases (MAPKs) cascades (ERK1/2, c-JNK, and p38), as well as nuclear translocation of NF-κB and AP-1 [[11], [12], [13]], which leads to release of multiple inflammatory mediators [[14], [15], [16]]. Furthermore, the activated MAPKs can phosphorylate some transcription factors to regulate target genes expression [17].

TRIM59 is a member of the tripartite motif-containing (TRIM) protein superfamily. It has a TRIM or RBCC motif consisting of a RING-finger domain (R), a B-box domain (B), and a coiled-coil domain [18,19]. Previous studies show that TRIM59 participates in many pathological processes, such as inflammation [5,20], cytotoxicity [21] and especially tumorigenesis [19,22]. TRIM59 interacts with evolutionarily conserved signaling intermediate in Toll pathways (ECSIT) and negatively regulates NF-κB and IRF-3/7-mediated signal pathways [23]. TRIM59 also regulates autophagy through modulating both transcription and ubiquitination of beclin 1 (BECN1) [24]. Moreover, TRIM59 plays a vital role in mouse early embryonic development stage, in which TRIM59 deficiency leads to embryonic lethality in mice by impairing gastrulation development [25]. Our previous study showed that TRIM59 expression was inhibited by LPS and TLR3 ligands in macrophages [5], but the underlying mechanisms remain to be elucidated.

Herein, we report that TRIM59 expression is controlled by Sp1 and Nrf1 in LPS-activated macrophages, which may be dependent on the activation of JNK signaling pathway. Our results also indicate that TRIM59 is highly expressed in macrophages, but significantly decreased in vivo and in vitro by LPS stimuli and during atherosclerosis progression. Combined with the anti-inflammatory properties of TRIM59 discovered in our previous work [5] and the function of inhibiting macrophage foam cells formation (data not shown), these results indicate that TRIM59 may be a potential therapeutic target for inflammatory diseases such as atherosclerosis.

Section snippets

Materials

LPS (L3129) and Wedelolactone (W4016) were purchased from Sigma. U0126 (HY-12031), SP600125 (HY-12041), SB203580 (HY-10256) and LY294002 (HY-10108) were obtained from MCE. Anti-TRIM59 (HPA017750, Sigma), Sp1 (CSB-PA004135, CusAb), Nrf1 (8052, CST, for Western blot; 12,936–1-AP, Proteintech, for CHIP array), TBP (ab818, Abcam) and Tubulin (sc-5286, Santa Cruz) antibodies were purchased as indicated.

Cell culture

RAW264.7 cells, a murine macrophage cell line, were purchased from Cell Bank of Chinese Academy of

Tissue and cellular distribution of TRIM59

To define the tissue distribution of TRIM59 in vivo, we extracted total proteins and RNA from mouse tissues and examined the TRIM59 expression by Western blot and qPCR. As shown in Fig. 1A, TRIM59 is highly expressed in liver, kidney, adipose, testis and lung, and moderately expressed in spleen, lymph node, thymus and aorta, but lowly expressed in brain, heart, muscle and bone marrow.

Next, we examined TRIM59 expression in macrophages, endothelial cells and smooth muscle cells because all these

Discussion

Macrophages play a critical role in the development of atherosclerosis. Although inhibiting macrophage functions has been well recognized as a therapeutic strategy in atherosclerosis, the therapeutic targets are still elusive. Recently, the importance of TRIM59 in macrophages has been recognized, but the regulatory mechanism for its expression remains unclear. Our previous findings [5] on the functions of TRIM59 in anti-inflammation and its down-regulation by LPS in macrophages require further

Author contributions

Yanying An, Zhihao Xu and Jiashu He conducted biochemical and in vivo experiments. Yuqi Ni, Shuizhen Shi and Yu Liu conducted molecular biology experiments. Ke-Yu Deng, Mingui Fu and Hong-Bo Xin edited the paper before submission. Meixiu Jiang designed experiments and prepared the paper.

Funding

This work was supported by National Natural Science Foundation of China grants (81760089 to MXJ and 81873658, 91639106 to HBX, 81760140, 8197021747 to KYD) and the grant for Jiangxi Provincial Collaborative Innovation Center of Biopharmaceutics and Biotechnology (2015202004 to HBX).

Declaration of Competing Interest

The Authors declare that there are no competing interests associated with the manuscript.

Acknowledgements

We thank Professor Yuanli Chen (Hefei University of Technology, College of Biomedical Engineering, China) for providing us the vector plasmid of pGL4.1 and pGL4.7 and his generous help.

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