Screening and bioinformatics analysis of cellular proteins interacting with chicken bromodomain-containing protein 2 in DF-1 cells.
Screening and bioinformatics analysis of cellular proteins interacting with chicken bromodomain-containing protein 2 in DF-1 cells.
复制标题
DF-1细胞中与鸡含溴结构域蛋白2相互作用的细胞蛋白的筛选和生物信息学分析。
DOI：
10.1080/00071668.2021.1943311
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发表时间：
2021
影响因子：
2
通讯作者：
Duan Zhiqiang
中科院分区：
文献类型：
--
作者：
Zhou Lei;Han Yifan;Yuan Cchao;Duan Zhiqiang
1.含溴结构域蛋白2（Bromodomain-containing protein 2，BRD 2）是BET蛋白家族的重要成员，能特异性结合组蛋白乙酰化赖氨酸参与基因转录调控、染色质重塑、细胞增殖和凋亡。对与chBRD 2相互作用的细胞蛋白的研究将有助于进一步了解chBRD 2的新功能和NDV的复制机制.将重组真核表达载体pEGFP-chBRD 2和空载体pEGFP-C1转染DF-1细胞，分别过表达GFP和GFP。利用GO注释、KEGG通路和蛋白质相互作用网络分析与chBRD 2相互作用的细胞蛋白。另外，选择一个靶蛋白，利用荧光共定位和Co-IP验证其与chBRD 2的相互作用。共鉴定了225种可能与chBRD 2相互作用的细胞蛋白。GO分析表明，这些在基因转录调控、细胞周期和发育、免疫和病毒感染中起着关键作用。进一步的KEGG通路分析表明，这些蛋白主要参与遗传信息处理、免疫系统、细胞过程和翻译。此外，还利用荧光共定位和Co-IP分析鉴定了一种靶向细胞蛋白鸡母蛋白3（chMATR 3）为chBRD 2复合物.本研究提供了chBRD 2蛋白在DF-1细胞中的相互作用组数据，为了解chBRD 2蛋白的功能提供了新的信息，为进一步研究NDV的复制和致病机制提供了新的靶点。
1. Bromodomain-containing protein 2 (BRD2) is an important member of the BET protein family, which can specifically bind histone acetylated lysine to participate in gene transcriptional regulation, chromatin remodelling, cell proliferation and apoptosis. The following investigation of cellular proteins interacting with chBRD2 will be helpful in understanding the new functions of chBRD2 and the mechanism of NDV replication.2. The recombinant eukaryotic expression vector pEGFP-chBRD2 and empty vector pEGFP-C1 were transfected into DF-1 cells to overexpress GFP-chBRD2 and GFP, respectively. GO annotation, KEGG pathway, and protein-protein interaction network were used to analyse the cellular proteins interacting with chBRD2. In addition, one targeted protein was selected to verify its interaction with chBRD2 using fluorescent co-localisation and Co-IP.3. A total of 225 cellular proteins were identified that potentially interact with chBRD2. GO analysis showed that these play key roles in gene transcriptional regulation, cell cycle and development, immunity and viral infection. Further KEGG pathway analysis showed that these proteins were mainly involved in genetic information processing, immune system, cellular processes and translation. In addition, one targeted cellular protein chicken matrin 3 (chMATR3) was also identified as chBRD2 complex using both fluorescence co-localisation and Co-IP analysis.4. This study presents the interactome data of chBRD2 protein in DF-1 cells, which provides new information to understand the functions of chBRD2 and new targets for further investigating the replication and pathogenesis of NDV.