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阿拉比多普西斯塔利亚纳植物染色相互作用因子3 (AtPIF3) bHLH和G-box之间的特定结合起源于胚胎细胞出现之前

Kuan-Ting Hsin¹, Yu-Hsuan Lee², Kai-Chun Lin³

¹Department of Tropical Agriculture and International Cooperation, National Pingtung University of Science and Technology, Pingtung, 912301, Taiwan.

BMC plant biology

|November 10, 2024

概括

基本螺旋环螺旋 (bHLH) 蛋白通过关键残留物结合DNA,G-box偏好在陆地植物 (胚胎细胞) 早期演变. 这项研究揭示了对植物发育至关重要的保存的DNA结合机制.

科学领域:

植物分子生物学植物分子生物学
进化生物学进化生物学
遗传学遗传学是一个

背景情况:

基本螺旋环螺旋 (bHLH) 蛋白质是植物发育至关重要的转录因子.
bHLH蛋白通过其基本区域的关键氨基酸残留物结合称为E盒 (5'-CANNTG-3') 的特定DNA序列.
对于E-box类型的精确选择以及这些DNA结合残留物在陆地植物 (胚胎细胞) 中bHLH蛋白中的进化保存,我们仍然不完全理解.

研究的目的:

调查关键氨基酸残留在阿拉比多普西斯塔利亚纳PIF3-bHLH (AtPIF3-bHLH) 的DNA结合特异性中的作用.
确定这些DNA识别位点在整个胚胎生物进化过程中的进化保护和功能意义.
阐明AtPIF3-bHLH的首选E盒类型和结合动态.

主要方法:

合成和分析复合的阿拉比多普西斯塔利亚纳PIF3-bHLH (AtPIF3-bHLH) 蛋白及其突变.
基于光的电泳移动性转移试验 (fEMSA) 来评估DNA结合能力和亲和力.
异热定位热量计 (ITC) 用于量化结合相互作用.
竞争性fEMSAs来确定电子盒的偏好.
分析净化选择信号和跨物种的DNA识别位置保护.

主要成果:

关键DNA识别部位的点突变显著降低了AtPIF3-bHLH的DNA结合能力和亲和力.
净化选择信号和DNA识别点的保护表明在整个胚胎生物进化过程中具有不变的作用.
在竞争性结合试验中,G-box优先被AtPIF3-bHLH结合,而不是其他E-box类型.
在AtPIF3-bHLH和G-box核心之间的动态结表明了核心区域的灵活识别.

结论:

关键氨基酸残留对于维持AtPIF3-bHLH-DNA相互作用至关重要.
在胚胎细胞进化的早期,bHLH-G-box识别机制被快速修复,突出了保留的角色.
这项研究为了解其他转录因子家族中的DNA识别决定因素提供了一个框架.

关键词:

循环节的时钟是循环节的时间.保护保护保护保护首选的DNA结合方式蛋白质-DNA相互作用

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