Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in <i>Apostichopus japonicus</i> following <i>Vibrio splendidus</i> infection

Lian-Lian Sun; Ying-Fen Dai; Mei-Xiang You; Cheng-Hua Li

doi:10.24272/j.issn.2095-8137.2023.106

Volume 44 Issue 5

Sep. 2023

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Lian-Lian Sun, Ying-Fen Dai, Mei-Xiang You, Cheng-Hua Li. Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in Apostichopus japonicus following Vibrio splendidus infection. Zoological Research, 2023, 44(5): 905-918. doi: 10.24272/j.issn.2095-8137.2023.106

Citation:

Lian-Lian Sun, Ying-Fen Dai, Mei-Xiang You, Cheng-Hua Li. Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in Apostichopus japonicus following Vibrio splendidus infection. Zoological Research, 2023, 44(5): 905-918. doi: 10.24272/j.issn.2095-8137.2023.106

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Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in Apostichopus japonicus following Vibrio splendidus infection

doi: 10.24272/j.issn.2095-8137.2023.106

1.
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo, Zhejiang 315211, China
2.
Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, Shandong 266071, China

Supplementary data to this article can be found online.
The authors declare that they have no competing interests.
L.L.S. conducted the experiments, processed the data, and wrote the manuscript; Y.F.D. and C.H.L. participated in the experimental design and revised the manuscript. M.X.Y. and C.H.L. contributed new reagents and analytic tools. All authors read and approved the final version of the manuscript.

Funds: This work was supported by the National Natural Science Foundation of China (32102825) and Natural Science Foundation of Zhejiang Province (LQ22C190003)

More Information

Corresponding author: E-mail: lichenghua@nbu.edu.cn
Received Date: 2023-05-21
Accepted Date: 2023-08-14

Published Online: 2023-08-14

Publish Date: 2023-09-18

Abstract

Abstract

Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in Apostichopus japonicus coelomocytes, leading to the production of excessive reactive oxygen species (ROS) and irreversible apoptotic cell death. Emerging evidence suggests that mitochondrial autophagy (mitophagy) is the most effective method for eliminating damaged mitochondria and ROS, with choline dehydrogenase (CHDH) identified as a novel mitophagy receptor that can recognize non-ubiquitin damage signals and microtubule-associated protein 1 light chain 3 (LC3) in vertebrates. However, the functional role of CHDH in invertebrates is largely unknown. In this study, we observed a significant increase in the mRNA and protein expression levels of A. japonicus CHDH (AjCHDH) in response to V. splendidus infection and lipopolysaccharide (LPS) challenge, consistent with changes in mitophagy under the same conditions. Notably, AjCHDH was localized to the mitochondria rather than the cytosol following V. splendidus infection. Moreover, AjCHDH knockdown using siRNA transfection significantly reduced mitophagy levels, as observed through transmission electron microscopy and confocal microscopy. Further investigation into the molecular mechanisms underlying CHDH-regulated mitophagy showed that AjCHDH lacked an LC3-interacting region (LIR) for direct binding to LC3 but possessed a FB1 structural domain that binds to SQSTM1. The interaction between AjCHDH and SQSTM1 was further confirmed by immunoprecipitation analysis. Furthermore, laser confocal microscopy indicated that SQSTM1 and LC3 were recruited by AjCHDH in coelomocytes and HEK293T cells. In contrast, AjCHDH interference hindered SQSTM1 and LC3 recruitment to the mitochondria, a critical step in damaged mitochondrial degradation. Thus, AjCHDH interference led to a significant increase in both mitochondrial and intracellular ROS, followed by increased apoptosis and decreased coelomocyte survival. Collectively, these findings indicate that AjCHDH-mediated mitophagy plays a crucial role in coelomocyte survival in A. japonicus following V. splendidus infection.
- Choline dehydrogenase,
- Mitophagy,
- SQSTM1,
- Microtubule-associated protein 1 light chain 3,
- Apostichopus japonicus

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References(60)

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