Page 116 - 《水产学报》2025年第11期

夏宁，等 水产学报, 2025, 49(11): 119409

Ubiquitination and phosphorylation modifications mediate the
macrophage lipid metabolism elicited by poly(I:C)

1,2
XIA Ning , SU Jianguo 1,2*
1. College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China;
2. Laboratory for Marine Biology and Biotechnology,
Qingdao Marine Science and Technology Center, Qingdao 266237, China

Abstract: Lipid droplets (LDs) are important organelles in the viral lifecycle, and viruses can regulate lipid metabolism and LD
synthesis through various mechanisms. Ubiquitination, phosphorylation, and glycosylation are classic protein modifications
known to participate in the regulation of host lipid metabolism. However, it remains unclear whether dsRNA viruses can
manipulate lipid metabolism through protein modifications. To investigate the roles of ubiquitination, phosphorylation, and N/O-
glycosylation modifications in the lipid metabolism process induced by dsRNA viruses, we conducted preliminary explorations
using proteomics and modificationomics techniques. Initially, primary kidney macrophages were isolated from Ctenopharyn-
godon idella using a percoll density gradient centrifugation and stimulated with poly(I:C) to simulate dsRNA virus infection.
Subsequently, differential proteins in the three modificationomics types were investigated, and GO function and KEGG path-
way enrichment were performed. The results revealed that 2 080 ubiquitinated proteins including 224 phosphorylation-modi-
fied differential proteins were identified in ubiquitination mass spectrometry. 8 lipid metabolism-related pathways encom-
passing 15 proteins were enriched in ubiquitination-modified differential proteins. Among all pathways enriched in ubiquitin
modification proteomics, lipid metabolism pathways were significantly enriched. Phosphorylation modification mass spectro-
metry detected 1 415 phosphorylated proteins, including 139 differential phosphorylated proteins. The phosphorylated differen-
tial proteins were enriched in only 1 lipid metabolism pathway and 1 significantly upregulated protein cytosolic phospholipase
A2 (cPLA2). N/O-glycosylation mass spectrometry analysis identified 371 N-glycosylated proteins and 243 O-glycosylated
proteins. Differential expression analysis revealed 368 N-glycosylated differential proteins and 120 O-glycosylated differential
proteins but no major lipid metabolism pathway was enriched among these differential proteins. Further, the expression levels
of the 16 differential modification proteins showed that ACOX1 and HADHA were the most significantly down-regulated pro-
teins in terms of ubiquitination modification following poly(I:C) stimulation, while DAGLα, ABCA1, and PLD1 were the most
significantly up-regulated proteins. The results suggested that ACOX1, HADHA, DAGLα, ABCA1, PLD1, and cPLA2 may be
the key modification proteins in the lipid metabolism induced by poly(I:C). In summary, the present study demonstrated that
ubiquitination and phosphorylation modifications play a crucial role in the lipid metabolism and lipid droplet synthesis in viral
dsRNA stimulation, underlying the important role of lipid metabolism-related protein modifications in viral dsRNA infections.
Key words: Ctenopharyngodon idella kidney macrophages; lipid metabolism; lipid droplets; poly(I:C); ubiquitination; phos-
phorylation; glycosylation

Corresponding author: SU Jianguo. E-mail: sujianguo@mail.hzau.edu.cn
Funding projects: National Natural Science Foundation of China (32373164)

https://www.china-fishery.cn 中国水产学会主办 sponsored by China Society of Fisheries
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