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在米中使用DREB1转录因子对DNA识别进行AlphaFold 3-Assisted解密

Wenshu Wang¹, Wei Cai¹, Jiang Zhu²

¹Institute of Crop Science of Wuhan Academy of Agricultural Sciences, Wuhan 430345, China.

International journal of molecular sciences

|July 12, 2025

概括

米的OsDREB1蛋白质对抗寒冷和干旱至关重要,通过它们的AP2域和KRP/RAGR基因结合DNA元素. 关键残留物R40和E66对于这种DNA识别机制至关重要.

科学领域:

植物分子生物学植物分子生物学
遗传学是一个遗传学.
生物化学生物化学

背景情况:

米的OsDREB1蛋白质是参与非生物应激耐受性 (寒冷,干旱) 的转录因子.
这些蛋白质与基因促进体中的特定DNA元素 (CRT,DRE,GCC-box) 结合.
OsDREB1s识别这些DNA元素的确切机制尚未完全理解.

研究的目的:

为了阐明大米OsDREB1蛋白的DNA识别机制.
研究特定氨基酸残留在DNA结合中的作用.
了解OsDREB1s如何与不同的DNA元素 (CRT,DRE,GCC-box) 相互作用.

主要方法:

使用AlphaFold进行蛋白质结构建模 3.
在模拟中,OsDREB1A与DNA元素结合.
电泳运动移位测试 (EMSA) 测定DNA结合亲和力.
位点定向突变发生,以验证关键残留物的功能.

主要成果:

在AlphaFold 3模型中,在OsDREB1s中发现了保守的AP2域和无序的KRP/RAGR动机.
预计关键残留物 (四个Arg,AP2中的E66,KRP/RAGR中的R40) 将与DNA主要和次要沟相互作用.
在EMSA的实验中,OsDREB1A与CRT,DRE和GCC-box的结合关系有所差异.
突变分析证实了R40和E66在CRT结合中的关键作用.

结论:

该研究提出了OsDREB1蛋白质的详细DNA识别机制,涉及AP2和KRP/RAGR领域的特定残留物.
这些发现提供了关于OsDREB1s如何与具有不同亲和力的不同DNA元素结合的见解.
鉴定出来的机制很可能在所有大米的OsDREB1蛋白中保留,这是由于保留的结构动机.

关键词:

AP2 域名域名AP2 域名域名 C-重复结合因子的重复无生物压力是无生物压力.结构建模结构建模转录法规转录法规转录法规

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