Study on the interaction protein of transcription factor Smad3 based on TurboID proximity labeling technology

IF 3.4 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Genomics Pub Date : 2024-03-26 DOI:10.1016/j.ygeno.2024.110839

Biao Yan , Ting Zeng , Xiaoshan Liu , Yuanyuan Guo , Hongguang Chen , Shuang Guo , Wu Liu

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引用次数: 0

Abstract

TurboID is a highly efficient biotin-labelling enzyme, which can be used to explore a number of new intercalating proteins due to the very transient binding and catalytic functions of many proteins. TGF-β/Smad3 signaling pathway is involved in many diseases, especially in diabetic nephropathy and inflammation. In this paper, a stably cell line transfected with Smad3 were constructed by using lentiviral infection. To further investigate the function of TGF-β/Smad3, the protein labeling experiment was conducted to find the interacting protein with Smad3 gene. Label-free mass spectrometry analysis was performed to obtain 491 interacting proteins, and the interacting protein hnRNPM was selected for IP and immunofluorescence verification, and it was verified that the Smad3 gene had a certain promoting effect on the expression of hnRNPM gene, and then had an inhibitory effect on IL-6. It lays a foundation for further study of the function of Smad3 gene and its involved regulatory network.

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基于 TurboID 近距离标记技术的转录因子 Smad3 互作蛋白研究

TurboID 是一种高效的生物素标记酶，由于许多蛋白质具有非常短暂的结合和催化功能，因此可用于探索许多新的插层蛋白。TGF-β/Smad3信号通路涉及多种疾病，尤其是糖尿病肾病和炎症。本文利用慢病毒感染技术构建了转染 Smad3 的稳定细胞系。为了进一步研究 TGF-β/Smad3 的功能，研究人员进行了蛋白标记实验，以寻找与 Smad3 基因相互作用的蛋白。通过无标记质谱分析得到491个相互作用蛋白，并选择相互作用蛋白hnRNPM进行IP和免疫荧光验证，验证了Smad3基因对hnRNPM基因的表达有一定的促进作用，进而对IL-6有抑制作用。这为进一步研究 Smad3 基因的功能及其参与的调控网络奠定了基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Genomics 生物-生物工程与应用微生物

CiteScore

9.60

自引率

2.30%

发文量

260

审稿时长

60 days

期刊介绍： Genomics is a forum for describing the development of genome-scale technologies and their application to all areas of biological investigation. As a journal that has evolved with the field that carries its name, Genomics focuses on the development and application of cutting-edge methods, addressing fundamental questions with potential interest to a wide audience. Our aim is to publish the highest quality research and to provide authors with rapid, fair and accurate review and publication of manuscripts falling within our scope.