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Functional analyses of TRAF6 gene in Argopecten scallops
Xiaotong Zhang1, Xia Wang1, Jieyu Chen2
1College of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, Shandong, 266109, China.
Fish & Shellfish Immunology
|February 14, 2024
Summary
Tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) plays a key role in the immune response of Argopecten scallops. TRAF6 knockdown significantly altered gene expression, revealing its involvement in essential cellular pathways for scallop immunity and breeding.
Area of Science:
- Marine Biology
- Immunology
- Genetics
Background:
- The tumor necrosis factor (TNF) receptor-associated factor (TRAF) family is crucial in immune signaling pathways.
- Previous work identified TRAF2, 3, 4, 6, and 7 in bay and Peruvian scallops.
- TRAF6 is a critical link in the TNF superfamily, prompting further investigation.
Purpose of the Study:
- To investigate the role and immune mechanism of TRAF6 in bay scallops (Argopecten irradians) and Peruvian scallops (Argopecten purpuratus) and their hybrids.
- To determine TRAF6's subcellular localization and its effect on NF-κB activity.
- To analyze the impact of TRAF6 knockdown on gene expression profiles in scallops.
Main Methods:
- Subcellular localization assays using HEK293T cells.
- Dual-luciferase reporter assays to assess NF-κB activation.
- RNA interference (RNAi) for TRAF6 knockdown.
- Transcriptomic analysis and KEGG enrichment analysis.
- Quantitative reverse transcription PCR (qRT-PCR) for gene expression validation.
Main Results:
- TRAF6 proteins were localized in the cytoplasm of HEK293T cells.
- TRAF6 overexpression significantly activated NF-κB in a dose-dependent manner.
- TRAF6 knockdown resulted in substantial changes in gene expression, with 1194, 2403, and 1099 differentially expressed genes in bay scallops, hybrids (Aip), and reciprocal hybrids (Api), respectively.
- Enriched pathways included transport and catabolism, amino acid metabolism, peroxisome, lysosome, and phagosome.
Conclusions:
- TRAF6 is essential for the immune response in Argopecten scallops.
- TRAF6 influences key cellular pathways involved in immunity.
- Understanding TRAF6 function can aid in scallop breeding programs.
