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转录阻遏物 VAL1 与转录共激活因子 SIN3 样蛋白(SNLs)之间的相互作用分析。

Interaction Analysis between the Transcription Repressor VAL1 and Transcription Coregulators SIN3-LIKEs (SNLs).

机构信息

State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, China.

Hubei Collaborative Innovation Center for Grain Industry, School of Agriculture, Yangtze University, Jingzhou 434023, China.

出版信息

Int J Mol Sci. 2022 Jun 23;23(13):6987. doi: 10.3390/ijms23136987.

Abstract

() encodes a DNA-binding B3 domain protein and plays essential roles in seed maturation and flowering transition by repressing genes through epigenetic silencing in . ()- (), which encode scaffold proteins for the assembly of histone deacetylase complexes and have six SIN3 homologues () in , directly repress gene expression to regulate seed maturation and flowering transition. However, it remains unclear whether VAL1 and SNLs work together in repressing the expression of related genes. In this study, yeast two-hybrid and firefly luciferase complementation imaging assays revealed that VAL1 interacts with SNLs, which can be attributed to its own zinc-finger CW (conserved Cys (C) and Trp (W) residues) domain and the PAH (Paired Amphipathic Helices) domains of SNLs. Furthermore, pull-down experiments confirmed that the CW domain of VAL1 interacts with both intact protein and the PAH domains of SNLs proteins, and the co-immunoprecipitation assays also confirmed the interaction between VAL1 and SNLs. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that and were expressed in seedlings, and transient expression assays showed that VAL1 and SNLs were localized in the nucleus. Considered together, these results reveal that VAL1 physically interacts with SNLs both in vitro and in vivo, and suggest that VAL1 and SNLs may work together to repress the expression of genes related to seed maturation and flowering transition in .

摘要

() 编码一种 DNA 结合 B3 结构域蛋白,通过在 中通过表观遗传沉默来抑制基因,从而在种子成熟和开花过渡中发挥重要作用。() 和 (),它们编码组蛋白去乙酰化酶复合物组装的支架蛋白,在 中有六个 SIN3 同源物 (),直接抑制基因表达以调节种子成熟和开花过渡。然而,目前尚不清楚 VAL1 和 SNLs 是否共同抑制相关基因的表达。在这项研究中,酵母双杂交和萤火虫荧光素酶互补成像测定表明 VAL1 与 SNLs 相互作用,这归因于其自身的锌指 CW(保守 Cys (C) 和 Trp (W) 残基)结构域和 SNLs 的 PAH(配对两性螺旋)结构域。此外,下拉实验证实 VAL1 的 CW 结构域与 SNLs 蛋白的完整蛋白和 PAH 结构域相互作用,共免疫沉淀实验也证实了 VAL1 和 SNLs 之间的相互作用。此外,定量实时 PCR (qRT-PCR) 分析显示 和 在幼苗中表达,瞬时表达实验表明 VAL1 和 SNLs 定位于细胞核中。综合这些结果表明,VAL1 与 SNLs 在体内和体外均具有物理相互作用,并且表明 VAL1 和 SNLs 可能共同作用以抑制与种子成熟和开花过渡相关的基因在 中的表达。

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