School of Life Science and Food Engineering, Hanshan Normal University, Chaozhou, 521041, China; Guangdong Provincial Key Laboratory of Functional Substances in Medicinal Edible Resources and Healthcare Products, Chaozhou, 521041, China.
Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou, 515063, China.
Dev Comp Immunol. 2024 Feb;151:105087. doi: 10.1016/j.dci.2023.105087. Epub 2023 Oct 26.
Hemocyanin is a respiratory protein, it is also a multifunctional immune molecule that plays a vital role against pathogen invasion in shrimp. However, the regulation of hemocyanin gene expression in shrimp hemocytes and the mechanisms involved during pathogen infection remains unclear. Here, we used DNA pull-down followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the Yin Yang 1 transcription factor homolog in Penaeus vannamei (PvYY1) as a key factor that modulates transcription of the small subunit hemocyanin gene of P. vannamei (PvHMCs) in hemocytes during Vibrio parahaemolyticus AHPND (VP) infection. Bioinformatics analysis revealed that the core promoter region of PvHMCs contains two YY1 motifs. Mutational and oligoprecipitation analyses confirmed that PvYY1 could bind to the YY1 motifs in the PvHMCs core promoter region, while truncation of PvYY1 revealed that the N-terminal domain of PvYY1 is essential for the transactivation of PvHMCs core promoter. Besides, the REPO domain of PvYY1 could repress the activity of the PvHMCs core promoter. Overexpression of PvYY1 significantly activates the promoter activity of PvHMCs core promoter, while PvYY1 knockdown significantly decreases the expression level of PvHMCs in shrimp hemocytes and survival rate of shrimp upon infection with VP. Our present study provides new insights into the transcriptional regulation of PvHMCs by PvYY1 in shrimp hemocytes during bacteria (VP) infection.
血蓝蛋白是一种呼吸蛋白,也是一种多功能免疫分子,在虾类抵御病原体入侵方面发挥着至关重要的作用。然而,虾血细胞中血蓝蛋白基因表达的调控及其在病原体感染过程中的机制仍不清楚。在这里,我们使用 DNA 下拉结合液相色谱-串联质谱(LC-MS/MS)鉴定了凡纳滨对虾中的 Yin Yang 1 转录因子同源物(PvYY1),它是一种关键因子,可调节在副溶血弧菌 AHPND(VP)感染期间虾血细胞中小亚基血蓝蛋白基因(PvHMCs)的转录。生物信息学分析表明,PvHMCs 的核心启动子区域包含两个 YY1 基序。突变和寡聚沉淀分析证实 PvYY1 可以与 PvHMCs 核心启动子区域的 YY1 基序结合,而 PvYY1 的截断分析表明 PvYY1 的 N 端结构域对于 PvHMCs 核心启动子的转录激活是必需的。此外,PvYY1 的 REPO 结构域可以抑制 PvHMCs 核心启动子的活性。过表达 PvYY1 可显著激活 PvHMCs 核心启动子的启动子活性,而 PvYY1 敲低则显著降低了虾血细胞中 PvHMCs 的表达水平和虾在感染 VP 后的存活率。本研究为副溶血弧菌感染期间 PvYY1 对虾血细胞中 PvHMCs 的转录调控提供了新的见解。