Screening of genes encoding proteins that interact with Nrf2: Probing a cDNA library from Mytilus coruscus using a yeast two-hybrid system
Screening of genes encoding proteins that interact with Nrf2: Probing a cDNA library from Mytilus coruscus using a yeast two-hybrid system
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DOI：
10.1016/j.fsi.2023.109112
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发表时间：
2023-09-24
影响因子：
4.7
通讯作者：
Qi，Pengzhi
中科院分区：
文献类型：
--
作者：
Zhu，Li;Li，Yaru;Qi，Pengzhi
核因子红细胞2相关因子2（Nrf 2）是生物体中最重要的内源性抗氧化因子，它通过直接或间接地与关键的先天免疫成分相互作用，对免疫应答发挥广泛的控制作用。虽然Nrf 2已被广泛证实参与哺乳动物和一些鱼类的抗应激性，但其对软体动物抗氧化应激性的贡献并不常见。本研究从中华绒螯蟹消化腺中提取总RNA。利用GATEWAY重组技术构建并筛选了coruscus的cDNA文库。克隆了M.将coruscus基因克隆到pGBKT 7载体中，制备诱饵质粒。利用酵母双杂交系统，经过营养缺陷型培养基筛选、测序和生物信息学分析，最终鉴定出13个与Nrf 2相互作用的结合蛋白。它们是QM样蛋白、40 S核糖体蛋白S4（RPS 4）、核糖体蛋白S2（RPS 2）、核糖体蛋白L12（RPL 12）、延伸因子1 α的EF 1-α mRNA（eEF 1-α）、铁蛋白、α-淀粉酶、胰蛋白酶、vdg 3、周期时钟蛋白、亲环蛋白A同种型1（cyclophilin A isoform 1，CRIA）、丝氨酸蛋白酶CFSP 2、组蛋白变体H2A.Z（H2A.Z）。为了更好地了解Nrf 2在动物中的生理功能，并作为未来研究Nrf 2相互作用中蛋白质作用的潜在目标，阐明这些蛋白质相互作用至关重要。
The Nuclear factor Erythroid 2-related factor 2 (Nrf2) is the most important endogenous antioxidant factor in organisms, and it has been demonstrated that it exerts extensive control over the immune response by interacting with crucial innate immunity components directly or indirectly. Although Nrf2 has been widely confirmed to be involved in stress resistance in mammals and some fish, its contribution to mollusks oxidative stress resistance has not frequently been documented. In this investigation, total RNA was taken from the digestive gland ofM. coruscus, and a cDNA library was constructed and screened using the GATEWAY recombination technology. The Nrf2 cDNA sequence ofM. coruscuswas cloned into the pGBKT7 vector to prepare the bait plasmid. Using yeast two-hybrid system, after auxotrophic medium screening, sequencing, and bioinformatics analysis, 13 binding proteins that interacted with Nrf2 were finally identified. They were QM-like protein, 40S ribosomal protein S4 (RPS4), ribosomal protein S2 (RPS2), ribosomal protein L12 (RPL12), EF1-alpha mRNA for elongation factor 1 alpha (eEF1-alpha), ferritin, alpha-amylase, trypsin, vdg3, period clock protein, cyclophilin A isoform 1 (CYP A), serine protease CFSP2, histone variant H2A.Z (H2A.Z). For a better understanding the physiological function of Nrf2 in animals and as a potential target for future research on protein roles in Nrf2 interactions, it is crucial to clarify these protein interactions.