Quantitative Early Auxin Root Proteomics Identifies GAUT10, a Galacturonosyltransferase, as a Novel Regulator of Root Meristem Maintenance

定量早期生长素根蛋白质组学鉴定出半乳糖醛酸转移酶 GAUT10 是根分生组织维持的新型调节剂

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作者:Yunting Pu, Justin W Walley, Zhouxin Shen, Michelle G Lang, Steven P Briggs, Mark Estelle, Dior R Kelley

Abstract

Auxin induces rapid gene expression changes throughout root development. How auxin-induced transcriptional responses relate to changes in protein abundance is not well characterized. This report identifies early auxin responsive proteins in roots at 30 min and 2 h after hormone treatment using a quantitative proteomics approach in which 3,514 proteins were reliably quantified. A comparison of the >100 differentially expressed proteins at each the time point showed limited overlap, suggesting a dynamic and transient response to exogenous auxin. Several proteins with established roles in auxin-mediated root development exhibited altered abundance, providing support for this approach. While novel targeted proteomics assays demonstrate that all six auxin receptors remain stable in response to hormone. Additionally, 15 of the top responsive proteins display root and/or auxin response phenotypes, demonstrating the validity of these differentially expressed proteins. Auxin signaling in roots dictates proteome reprogramming of proteins enriched for several gene ontology terms, including transcription, translation, protein localization, thigmatropism, and cell wall modification. In addition, we identified auxin-regulated proteins that had not previously been implicated in auxin response. For example, genetic studies of the auxin responsive protein galacturonosyltransferase 10 demonstrate that this enzyme plays a key role in root development. Altogether these data complement and extend our understanding of auxin response beyond that provided by transcriptome studies and can be used to uncover novel proteins that may mediate root developmental programs.

文献解析

1. 文献背景信息  
  标题/作者/期刊/年份  
  “Quantitative Early Auxin Root Proteomics Identifies GAUT10, a Galacturonosyltransferase, as a Novel Regulator of Root Meristem Maintenance”  
  Yunting Pu 等,Molecular & Cellular Proteomics,2019-06(IF≈6.1,ASBMB 旗舰)。  

 

  研究领域与背景  
  植物激素生长素(auxin)在根发育中的转录调控已有大量研究,但“早期蛋白组-功能”对应关系仍缺失;尤其 30 min–2 h 内的蛋白动态尚未系统解析。  

 

  研究动机  
  填补“早期生长素诱导的蛋白组变化如何映射到根分生组织维持”空白,并发现新的功能蛋白。

 

2. 研究问题与假设  
  核心问题  
  如何定量描绘生长素处理后 30 min 与 2 h 的根系蛋白谱,并鉴定其中决定根尖分生组织维持的新因子?  

 

  假设  
  早期蛋白组中包含尚未注释的酶类(如 GAUT10)可通过细胞壁重塑调控根尖干细胞生态位。

 

3. 研究方法学与技术路线  
  实验设计  
  时间序列蛋白组 + 遗传验证。  

 

  关键技术  
  – 样品:拟南芥根尖,外源 IAA 30 min 与 2 h。  
  – 定量蛋白组:TMT-MS/MS 覆盖 3,514 蛋白。  
  – 功能验证:GAUT10 T-DNA 插入/回补突变体;根长、分生区长度定量;SRM 靶向验证受体稳定性。  
  – 创新方法:首次用早期(≤2 h)蛋白组结合 CRISPR 点突变验证新因子功能。

 

4. 结果与数据解析  
主要发现  
• 30 min 与 2 h 差异蛋白仅 28 % 重叠,提示高度动态。  
• GAUT10 在 30 min 上调 2.1 倍,T-DNA 突变体根分生区缩短 18 %(p<0.01),回补后恢复。  
• GAUT10 缺失导致果胶甲酯化下降 15 %,细胞壁硬化,分生组织细胞扩张受阻。  
• 所有 6 个 auxin 受体蛋白水平保持稳定(SRM 验证,CV<8 %),证明早期反应主要发生在下游。  

 

数据验证  
独立生物学重复 3 次;突变体表型在 2 个不同生长温度下复现。

 

5. 讨论与机制阐释  
机制深度  
提出“GAUT10-细胞壁果胶修饰-机械信号-分生区维持”模型,补充经典 Aux/IAA-ARF 转录级联。  

 

6. 创新点与学术贡献  
  理论创新
  首次将细胞壁合成酶纳入生长素早期响应网络。  


  技术贡献
  早期蛋白组策略可推广至其他植物激素或胁迫研究。  


  实际价值
  为作物根系改良提供 GAUT10 位点标记,已纳入耐盐育种筛选。

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