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The interaction of O-GlcNAc-modified NLRX1 and IKK-α modulates IL-1β expression in M1 macrophages

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Abstract

NOD-like receptor (NLR)X1 (NLRX1) is a negative regulator of inflammation by inhibiting nuclear factor-κB (NF-κB) signaling and downstream pro-inflammatory factors. However, its post-translational modification and how it participates in regulating the inflammatory responses in macrophages are still unclear. Here, we found that NLRX1 was modified with O-linked N-acetylglucosamine (O-GlcNAc). The interaction and co-localization between NLRX1 and O-GlcNAc transferase (OGT) was validated by co-immunoprecipitation and confocal microscopy analysis, and the nucleotide-binding domain (NBD) region of NLRX1 was required for its interaction with OGT. NLRX1 protein increased significantly after treatment with a high dose of OGT inhibitor OSMI-1. Elevated O-GlcNAcylation level promoted NLRX1 ubiquitination and decreased NLRX1 stability proved by ubiquitination and cycloheximide (CHX) chase experiments, and enhanced the interaction between NLRX1 and inhibitor of nuclear factor kappaB kinase-α (IKK-α), thus reducing the expression of inflammatory cytokine IL-1β in M1 macrophages. Together, our results indicate that the interaction between NLRX1 and O-GlcNAcylation coordinates and modulates the inflammatory process in macrophages.

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Data availability

The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.

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Funding

The work was supported by the National Natural Science Foundation of China (Grant No. 81102850) and the Innovation and Entrepreneurship training Program for College Students of Guangdong Medical University (No. S202010571060)

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  1. Liqiong Chen and Yueliang Li are contributed equally

Authors and Affiliations

  1. Department of Clinical Immunology, Institute of Laboratory Medicine, Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China

    Liqiong Chen, Yueliang Li, Shuxian Zeng, Shujuan Duan, Zhuanglin Huang & Yi Liang

  2. Shenzhen Bao’an District Songgang People’s Hospital, 2 Shajiang Road, Bao’an District, Shenzhen, 518100, Guangdong, China

    Yueliang Li

  3. Clinical Laboratory, Liangzhu Hospital, 1657 Moganshan Road, Yuhang District, Hangzhou, 311100, Zhejiang, China

    Zhuanglin Huang

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  1. Liqiong Chen
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Contributions

LQC wrote the original manuscript; LQC and YLL performed and analyzed most of the experiments; SXZ and SJD conducted the experiments; ZLH performed data analysis; YL designed the experiment, and made critical revision and final approval of manuscript.

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Correspondence to Yi Liang.

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Chen, L., Li, Y., Zeng, S. et al. The interaction of O-GlcNAc-modified NLRX1 and IKK-α modulates IL-1β expression in M1 macrophages. In Vitro Cell.Dev.Biol.-Animal 58, 408–418 (2022). https://doi.org/10.1007/s11626-022-00654-1

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