Ubiquitylation of MFHAS1 by the ubiquitin ligase praja2 promotes M1 macrophage polarization by activating JNK and p38 pathways

Jing Zhong; Huihui Wang; Wankun Chen; Zhirong Sun; Jiawei Chen; Yajun Xu; Meilin Weng; Qiqing Shi; Duan Ma; Changhong Miao

doi:10.1038/cddis.2017.102

Ubiquitylation of MFHAS1 by the ubiquitin ligase praja2 promotes M1 macrophage polarization by activating JNK and p38 pathways

Cell Death Dis. 2017 May 4;8(5):e2763. doi: 10.1038/cddis.2017.102.

Authors

Jing Zhong^{1

2}, Huihui Wang³, Wankun Chen^{1

2}, Zhirong Sun^{1

2}, Jiawei Chen^{1

2}, Yajun Xu^{1

2}, Meilin Weng^{1

2}, Qiqing Shi⁴, Duan Ma⁵, Changhong Miao^{1

2

3}

Affiliations

¹ Department of Anesthesiology, Fudan University Shanghai Cancer Center, Shanghai, China.
² Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.
³ Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College, Xuzhou, China.
⁴ Department of Anesthesiology, Children's Hospital of Fudan University, Shanghai, China.
⁵ Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Collaborative Innovation Center of Genetics and Development, Institute of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai, China.

Abstract

Sepsis is a systemic inflammation caused by infection. The balance between M1-M2 macrophage polarization has an essential role in the pathogenesis of sepsis. However, the exact mechanism underlying macrophage polarization is unclear. We previously showed that levels of malignant fibrous histiocytoma amplified sequence 1 (MFHAS1) were significantly elevated in septic patients compared with those in nonseptic patients, and involved in the activation of Toll-like receptor (TLR) 2/c-Jun N-terminal kinase (JNK)/nuclear factor (NF)-κB pathway. In the present study, we explored whether MFHAS1 was involved in macrophage polarization and determined the effect of MFHAS1 on inflammation. We performed in vitro pulldown assays and in vivo co-immunoprecipitation assays and found that E3 ubiquitin ligase praja2 could directly bind to MFHAS1. In situ immunostaining analysis confirmed the colocalization of endogenous praja2 with MFHAS1. We first reported that praja2 promotes the accumulation of ubiquitylated MFHAS1 but does not degrade it. Moreover, our results indicate that MFHAS1 ubiquitylation by praja2 positively regulates TLR2-mediated JNK/p38 pathway and promotes M1 macrophage polarization, M2 to M1 macrophage transformation and inflammation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Cycle Proteins / chemistry
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism*
Cell Polarity / drug effects
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
HEK293 Cells
Humans
Imidazoles / pharmacology
Inflammation / metabolism
Inflammation / pathology
Interleukin-6 / genetics
Interleukin-6 / metabolism
JNK Mitogen-Activated Protein Kinases / metabolism*
Lipopeptides / pharmacology
Macrophages / cytology
Macrophages / drug effects
Macrophages / metabolism
Mice
Nitric Oxide Synthase Type II / genetics
Nitric Oxide Synthase Type II / metabolism
Oncogene Proteins / chemistry
Oncogene Proteins / genetics
Oncogene Proteins / metabolism*
Pyridines / pharmacology
RAW 264.7 Cells
Signal Transduction / drug effects
Toll-Like Receptor 2 / metabolism
Tumor Necrosis Factor-alpha / genetics
Tumor Necrosis Factor-alpha / metabolism
Ubiquitin-Protein Ligases / chemistry
Ubiquitin-Protein Ligases / metabolism*
Ubiquitination / drug effects
p38 Mitogen-Activated Protein Kinases / metabolism*

Substances

Cell Cycle Proteins
DNA-Binding Proteins
Imidazoles
Interleukin-6
Lipopeptides
MFHAS1 protein, human
Oncogene Proteins
Pam(3)CSK(4) peptide
Pyridines
Toll-Like Receptor 2
Tumor Necrosis Factor-alpha
Nitric Oxide Synthase Type II
PJA2 protein, human
Ubiquitin-Protein Ligases
JNK Mitogen-Activated Protein Kinases
p38 Mitogen-Activated Protein Kinases
SB 203580