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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...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9879056/ https://www.ncbi.nlm.nih.gov/pubmed/36713402 http://dx.doi.org/10.3389/fimmu.2022.1091419 |
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author | Liu, Wenjuan Ma, Jilv Chen, Jiwen Huang, Baoyu Liu, Fengchen Li, Lingling Fan, Nini Li, Fangshu Zheng, Yanxin Zhang, Xuekai Wang, Xiaona Wang, Xiaomei Wei, Lei Liu, Yaqiong Zhang, Meiwei Han, Yijing Wang, Xiaotong |
author_facet | Liu, Wenjuan Ma, Jilv Chen, Jiwen Huang, Baoyu Liu, Fengchen Li, Lingling Fan, Nini Li, Fangshu Zheng, Yanxin Zhang, Xuekai Wang, Xiaona Wang, Xiaomei Wei, Lei Liu, Yaqiong Zhang, Meiwei Han, Yijing Wang, Xiaotong |
author_sort | Liu, Wenjuan |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-9879056 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-98790562023-01-27 A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri Liu, Wenjuan Ma, Jilv Chen, Jiwen Huang, Baoyu Liu, Fengchen Li, Lingling Fan, Nini Li, Fangshu Zheng, Yanxin Zhang, Xuekai Wang, Xiaona Wang, Xiaomei Wei, Lei Liu, Yaqiong Zhang, Meiwei Han, Yijing Wang, Xiaotong Front Immunol Immunology 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. Frontiers Media S.A. 2023-01-12 /pmc/articles/PMC9879056/ /pubmed/36713402 http://dx.doi.org/10.3389/fimmu.2022.1091419 Text en Copyright © 2023 Liu, Ma, Chen, Huang, Liu, Li, Fan, Li, Zheng, Zhang, Wang, Wang, Wei, Liu, Zhang, Han and Wang https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Liu, Wenjuan Ma, Jilv Chen, Jiwen Huang, Baoyu Liu, Fengchen Li, Lingling Fan, Nini Li, Fangshu Zheng, Yanxin Zhang, Xuekai Wang, Xiaona Wang, Xiaomei Wei, Lei Liu, Yaqiong Zhang, Meiwei Han, Yijing Wang, Xiaotong A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri |
title | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
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title_full | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
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title_fullStr | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
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title_full_unstemmed | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
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title_short | A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri
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title_sort | novel tbk1/ikkϵ is involved in immune response and interacts with myd88 and mavs in the scallop chlamys farreri |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9879056/ https://www.ncbi.nlm.nih.gov/pubmed/36713402 http://dx.doi.org/10.3389/fimmu.2022.1091419 |
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