An HECT domain ubiquitin ligase CgWWP1 regulates granulocytes proliferation in oyster Crassostrea gigas. (October 2021)

• Record Type: Journal Article
• Authors: Song, Ying; Song, Xiaorui; Zhang, Dan; Yang, Ying; Wang, Lingling; Song, Linsheng
• Is Part Of: Developmental and comparative immunology. Volume 123(2021)
• Publication Date: 2021-10
• Subjects: Granulocytes -- Proliferation activity -- CgWWP1 -- Ubiquitination -- Granulocyte potential molecular markers; Immunology -- Periodicals; Developmental immunology -- Periodicals; 616.079
• DOI: 10.1016/j.dci.2021.104148
• Languages: English
• ISSNs: 0145-305X

Abstract

Abstract: Ubiquitination is involved in the regulation of granulocyte proliferation in vertebrate, and E3 ubiquitin ligase WWP1 has been reported to play an essential role in this process. In the present study, an HECT type E3 ubiquitin ligase ( Cg WWP1) was identified from oyster Crassostrea gigas, which contained a N-terminal C2 domain, four WW domains, and a C-terminal HECT domain. Cg WWP1 was able to bind the activated ubiquitin (Ub) and formed Cg WWP1-Ub complex in vitro . The mRNA transcripts of Cg WWP1 were expressed in granulocytes, semi-granulocytes and agranulocytes, with the highest expression level in granulocytes. The expressions of potential granulocyte markers Cg SOX11 (0.18-fold, p < 0.05) and Cg AATase (0.2-fold, p < 0.01) in haemocytes were significantly down-regulated at 24 h after the treatment with Indole-3-carbinol (I3C), a WWP1 inhibitor. The proportions of EdU + granulocytes reduced significantly at 12 h (8.1% ± 1.4%) and 24 h (9.7% ± 2.8%) after I3C treatment, which were significantly lower than that in the sterile seawater treatment (SW) group at 12 h (15.8% ± 4.2%) and 24 h (17.6% ± 0.8%), respectively. Meanwhile, the green EdU signals observed by confocal scanning microscopy in granulocytes of oysters treated by I3C became weaker compared to that in the SW group. These results indicated that Cg WWP1 was involved in the regulation of granulocyte proliferation as a ubiquitin-protein ligase. Highlights: A typical E3 ligase enzyme Cg WWP1 was identified from oyster. r Cg WWP1 was able to bind ubiquitin form to Cg WWP1-Ub complex in vitro . The inhibition of Cg WWP1 reduced the expression of Cg AATase and Cg SOX11. Cg WWP1 promoted granulocyte proliferation as a ubiquitin-protein ligase.