ABSTRACT

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.

摘要

1、含溴结构域蛋白2（BRD2）是BET蛋白家族的重要成员，可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。以下对与chBRD2相互作用的细胞蛋白的研究将有助于理解chBRD2的新功能和NDV复制机制。

2、将重组真核表达载体pEGFP-chBRD2和空载体pEGFP-C1转染DF-1细胞，分别过表达GFP-chBRD2和GFP。GO注释、KEGG通路和蛋白质-蛋白质相互作用网络用于分析与chBRD2相互作用的细胞蛋白质。此外，使用荧光共定位和 Co-IP 选择了一种靶向蛋白质来验证其与 chBRD2 的相互作用。

3. 共鉴定了 225 种可能与 chBRD2 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关键作用。进一步的KEGG通路分析表明，这些蛋白质主要参与遗传信息处理、免疫系统、细胞过程和翻译。此外，还使用荧光共定位和 Co-IP 分析将一种靶向细胞蛋白鸡 matrin 3 (chMATR3) 鉴定为 chBRD2 复合物。

4. 本研究提供了 DF-1 细胞中 chBRD2 蛋白的相互作用组数据，为了解 chBRD2 的功能提供了新的信息，为进一步研究 NDV 的复制和发病机制提供了新的靶点。