A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
Inhibitor of κB kinase (IKK) family proteins are key signaling molecules in the animal innate immune system and are considered master regulators of inflammation and innate immunity that act by controlling the activation of transcription factors such as NF-κB. However, few functional studies on IKK i...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1091419/full |
| _version_ | 1797954807921639424 |
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| author | Wenjuan Liu Jilv Ma Jiwen Chen Baoyu Huang Fengchen Liu Lingling Li Lingling Li Nini Fan Fangshu Li Yanxin Zheng Xuekai Zhang Xiaona Wang Xiaomei Wang Lei Wei Yaqiong Liu Meiwei Zhang Yijing Han Xiaotong Wang |
| author_facet | Wenjuan Liu Jilv Ma Jiwen Chen Baoyu Huang Fengchen Liu Lingling Li Lingling Li Nini Fan Fangshu Li Yanxin Zheng Xuekai Zhang Xiaona Wang Xiaomei Wang Lei Wei Yaqiong Liu Meiwei Zhang Yijing Han Xiaotong Wang |
| author_sort | Wenjuan Liu |
| collection | DOAJ |
| description | Inhibitor of κB kinase (IKK) family proteins are key signaling molecules in the animal innate immune system and are considered master regulators of inflammation and innate immunity that act by controlling the activation of transcription factors such as NF-κB. However, few functional studies on IKK in invertebrates have been conducted, especially in marine mollusks. In this study, we cloned the IKK gene in the Zhikong scallop Chlamys farreri and named it CfIKK3. CfIKK3 encodes a 773-amino acid-long protein, and phylogenetic analysis showed that CfIKK3 belongs to the invertebrate TBK1/IKKϵ protein family. Quantitative real-time PCR analysis showed that CfIKK3 mRNA is ubiquitously expressed in all tested scallop tissues. The expression of CfIKK3 transcripts was significantly induced after challenge with lipopolysaccharide, peptidoglycan, or poly(I:C). Co-immunoprecipitation (co-IP) assays confirmed the direct interaction of CfIKK3 with MyD88 (the key adaptor in the TLR pathway) and MAVS (the key adaptor in the RLR pathway), suggesting that this IKK protein plays a crucial role in scallop innate immune signal transduction. In addition, the CfIKK3 protein formed homodimers and bound to CfIKK2, which may be a key step in the activation of its own and downstream transcription factors. Finally, in HEK293T cells, dual-luciferase reporter gene experiments showed that overexpression of CfIKK3 protein activated the NF-κB reporter gene in a dose-dependent manner. In conclusion, our experimental results confirmed that CfIKK3 could respond to PAMPs challenge and participate in scallop TLR and RLR pathway signaling, ultimately activating NF-κB. Therefore, as a key signaling molecule and modulator of immune activity, CfIKK3 plays an important role in the innate immune system of scallops. |
| first_indexed | 2024-04-10T23:24:22Z |
| format | Article |
| id | doaj.art-9e958ba8df9246d79e00f309a4990022 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-10T23:24:22Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj.art-9e958ba8df9246d79e00f309a49900222023-01-12T12:53:06ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-01-011310.3389/fimmu.2022.10914191091419A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreriWenjuan Liu0Jilv Ma1Jiwen Chen2Baoyu Huang3Fengchen Liu4Lingling Li5Lingling Li6Nini Fan7Fangshu Li8Yanxin Zheng9Xuekai Zhang10Xiaona Wang11Xiaomei Wang12Lei Wei13Yaqiong Liu14Meiwei Zhang15Yijing Han16Xiaotong Wang17School of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaOcean School, Yantai University, Yantai, ChinaChangdao Enhancement and Experiment Station, Chinese Academy of Fishery Sciences, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaChangdao Enhancement and Experiment Station, Chinese Academy of Fishery Sciences, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaChangdao Enhancement and Experiment Station, Chinese Academy of Fishery Sciences, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaSchool of Agriculture, Ludong University, Yantai, ChinaInhibitor of κB kinase (IKK) family proteins are key signaling molecules in the animal innate immune system and are considered master regulators of inflammation and innate immunity that act by controlling the activation of transcription factors such as NF-κB. However, few functional studies on IKK in invertebrates have been conducted, especially in marine mollusks. In this study, we cloned the IKK gene in the Zhikong scallop Chlamys farreri and named it CfIKK3. CfIKK3 encodes a 773-amino acid-long protein, and phylogenetic analysis showed that CfIKK3 belongs to the invertebrate TBK1/IKKϵ protein family. Quantitative real-time PCR analysis showed that CfIKK3 mRNA is ubiquitously expressed in all tested scallop tissues. The expression of CfIKK3 transcripts was significantly induced after challenge with lipopolysaccharide, peptidoglycan, or poly(I:C). Co-immunoprecipitation (co-IP) assays confirmed the direct interaction of CfIKK3 with MyD88 (the key adaptor in the TLR pathway) and MAVS (the key adaptor in the RLR pathway), suggesting that this IKK protein plays a crucial role in scallop innate immune signal transduction. In addition, the CfIKK3 protein formed homodimers and bound to CfIKK2, which may be a key step in the activation of its own and downstream transcription factors. Finally, in HEK293T cells, dual-luciferase reporter gene experiments showed that overexpression of CfIKK3 protein activated the NF-κB reporter gene in a dose-dependent manner. In conclusion, our experimental results confirmed that CfIKK3 could respond to PAMPs challenge and participate in scallop TLR and RLR pathway signaling, ultimately activating NF-κB. Therefore, as a key signaling molecule and modulator of immune activity, CfIKK3 plays an important role in the innate immune system of scallops.https://www.frontiersin.org/articles/10.3389/fimmu.2022.1091419/fullChlamys farreriinnate immunityTBK1/IKKϵMyD88MAVS |
| spellingShingle | Wenjuan Liu Jilv Ma Jiwen Chen Baoyu Huang Fengchen Liu Lingling Li Lingling Li Nini Fan Fangshu Li Yanxin Zheng Xuekai Zhang Xiaona Wang Xiaomei Wang Lei Wei Yaqiong Liu Meiwei Zhang Yijing Han Xiaotong Wang A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri Frontiers in Immunology Chlamys farreri innate immunity TBK1/IKKϵ MyD88 MAVS |
| title | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
| title_full | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
| title_fullStr | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
| title_full_unstemmed | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
| title_short | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
| title_sort | novel tbk1 ikkϵ is involved in immune response and interacts with myd88 and mavs in the scallop chlamys farreri |
| topic | Chlamys farreri innate immunity TBK1/IKKϵ MyD88 MAVS |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1091419/full |
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