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钙调素样蛋白 CML43 作为水杨酸诱导的根特异性 Ca(2+) 传感器在拟南芥中发挥作用。

The calmodulin-like protein CML43 functions as a salicylic-acid-inducible root-specific Ca(2+) sensor in Arabidopsis.

机构信息

*Department of Biology, Queen's University, Kingston, ON, Canada, K7L 3N6.

出版信息

Biochem J. 2014 Jan 1;457(1):127-36. doi: 10.1042/BJ20131080.

Abstract

Many signalling pathways in plants are regulated by the second messenger calcium (Ca(2+)). In the standard model, Ca(2+)-sensor proteins, such as CaM (calmodulin), detect Ca(2+) signals and subsequently regulate downstream targets to advance the signal transduction cascade. In addition to CaM, plants possess many CMLs (CaM-like proteins) that are predicted to function as Ca(2+) sensors, but which remain largely uncharacterized. In the present study, we examined the biochemical properties, subcellular localization and tissue-specific distribution of Arabidopsis CML43. Our data indicate that CML43 displays characteristics typical of Ca(2+) sensors, including high-affinity Ca(2+) binding, conformational changes upon Ca(2+) binding that expose hydrophobic regions and stabilization of structure in the presence of Mg(2+) or Ca(2+). In vivo localization analysis demonstrates that CML43 resides in cytosolic and nuclear compartments. Transgenic plants expressing a CML43:GUS (β-glucoronidase) promoter reporter gene revealed that CML43 promoter activity is restricted almost exclusively to root tips under normal growth conditions. GUS reporter activity in these transgenic plants was strongly increased when exposed to the defence compound SA (salicylic acid). Furthermore, immunoblot analysis revealed that the CML43 protein accumulates following treatment with SA. Collectively, our findings suggest that CML43 functions as a Ca(2+) sensor in root tips during both normal growth and plant immune response.

摘要

许多植物中的信号通路都受到第二信使钙(Ca(2+))的调节。在标准模型中,Ca(2+)传感器蛋白,如 CaM(钙调蛋白),检测 Ca(2+)信号,然后调节下游靶标以推进信号转导级联。除了 CaM,植物还拥有许多 CML(CaM 样蛋白),这些蛋白被预测作为 Ca(2+)传感器发挥作用,但它们在很大程度上仍未被描述。在本研究中,我们研究了拟南芥 CML43 的生化特性、亚细胞定位和组织特异性分布。我们的数据表明,CML43 显示出典型的 Ca(2+)传感器特征,包括高亲和力的 Ca(2+)结合、Ca(2+)结合后暴露疏水区的构象变化以及在 Mg(2+)或 Ca(2+)存在下稳定结构。体内定位分析表明 CML43 存在于细胞质和核区室中。表达 CML43:GUS(β-葡糖苷酸酶)启动子报告基因的转基因植物表明,在正常生长条件下,CML43 启动子活性几乎仅局限于根尖。在这些转基因植物中,当暴露于防御化合物 SA(水杨酸)时,GUS 报告基因活性强烈增加。此外,免疫印迹分析表明,在 SA 处理后 CML43 蛋白积累。总的来说,我们的发现表明 CML43 在正常生长和植物免疫反应期间作为 Ca(2+)传感器在根尖中发挥作用。

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