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IKKε-like plays an important role in the innate immune signaling of the Pacific oyster (Crassostrea gigas)

Author:
Huang, Baoyu, Tang, Xueying, Zhang, Linlin, Li, Li, Wang, Wei, Liu, Mingkun, Zhang, Guofan
Source:
Fish & shellfish immunology 2019 v.93 pp. 551-558
ISSN:
1050-4648
Subject:
Crassostrea gigas, amino acids, complementary DNA, enzymes, gene activation, gene expression, hemocytes, human cell lines, immune system, interferons, messenger RNA, mutants, open reading frames, oysters, phosphorylation, polyinosinic-polycytidylic acid, quantitative polymerase chain reaction, reporter genes, shellfish, signal transduction, transcription factor NF-kappa B, transfection
Abstract:
IκB-related kinase ε (IKKε) plays a crucial role in the activation of nuclear factor κB (NF-κB) by phosphorylating inhibitor of NF-κB (IκB) and in the regulation of interferon (IFN) gene expression by phosphorylating IFN regulatory factors (IRFs). In this study, we cloned an IKKε homologue cDNA (designated as CgIKKε-like) from the Pacific oyster, Crassostrea gigas. The full 2896-bp cDNA sequence comprised a 2163-bp open reading frame (ORF) encoding 720 amino acids. CgIKKε-like is ubiquitously expressed, and its mRNA levels in hemocytes after poly I:C, V. alginolyticus, or OsHV-1 μVar challenge were analyzed by real-time PCR. Compared to that in the control, CgIKKε-like mRNA expression levels were significantly increased at 3 h and peaked at 6 h after OsHV-1 μVar challenge; no obvious changes were observed in expression levels until 24 h after either V. alginolyticus or poly I:C challenge, reaching a maximum at 24 h (p < 0.01) and then rapidly decreasing. CgIKKε-like transfection into human cell lines induced NF-κB and ISRE activation, while transfection with CgIKKε-like deletion mutants abolished NF-κB and ISRE reporter gene activation. Additionally, CgIKKε-like could interact with CgTBK1 and could form homodimers strongly, which may be critical for the immune signaling transduction. Last but not least, we found that CgIKKε-like may increase CgIκBs phosphorylation and could interact with CgIRF8. Together, these results suggest that CgIKKε-like could respond to pathogenic infection, participate in the immune signal transduction and activate NF-κB and ISRE reporter genes. Thus, CgIKKε-like could play an important role in the oyster immune system.
Agid:
6553996