首席研究员Plant Cell揭示多肽受体蛋白相互作用机制

【字体: 时间:2010年03月25日 来源:生物通

编辑推荐:

  中国科学院植物研究所分子发育生物学研究中心林金星研究员近期运用萤火虫荧光素酶互补技术揭示多肽激素CLV3受体蛋白之间的相互作用,相关成果文章Analysis of interactions among the CLAVATA3 receptors reveals a direct interaction between CLAVATA2 and CORYNE in Arabidopsis公布在Plant Journal上。

  

生物通报道,中国科学院植物研究所分子发育生物学研究中心林金星研究员近期运用萤火虫荧光素酶互补技术揭示多肽激素CLV3受体蛋白之间的相互作用,相关成果文章Analysis of interactions among the CLAVATA3 receptors reveals a direct interaction between CLAVATA2 and CORYNE in Arabidopsis公布在Plant Journal上。

 

文章通讯作者是林金星首席研究员,现任植物分子与发育生物学研究中心主任,主要从事植物细胞与生殖生物学方面的研究。

 

高等植物多肽激素CLAVATA3(CLV3)对于植物茎端分生组织干细胞数目的维持起着极其重要的作用。过去几十年来,CLAVATA1/CLAVATA2 (CLV1/CLV2) 复合体被认为是CLV3多肽在细胞膜上的唯一受体。

 

然而最近的遗传学分析筛选到了一个新的受体蛋白激酶成员—CORYNE(CRN),并且发现它在CLV3信号途径中起着非常重要的作用。在一系列遗传学分析的基础上,新的CLV3信号转导通路假设被提出:即CLV1同源二聚体与CLV2/CRN异源复合体可能平行独立地介导CLV3信号。

 

然而,新的假设仅仅停留在遗传学上的预测,仍然缺乏进一步的生物化学和细胞学的证据。为了更好地理解这三个可能的受体蛋白之间的相互关系,林金星研究组利用新型的活体下检测蛋白质相互作用的技术—萤火虫荧光素酶互补技术(Firefly luciferase complementation imaging assay, LCI) 在拟南芥叶肉原生质体 (Arabidopsis mesophyll protoplasts) 和烟草叶片 (Nicotiana benthamiana leaves) 两个体系中分析了CLV1, CLV2CRN三个受体蛋白之间的相互作用。

 

实验结果表明:LCI技术和免疫共沉淀技术 (Co-immunoprecipitation assay) 都证实了CLV2在没有CLV3多肽刺激的情况下可以直接地与CRN发生相互作用;外源的CLV3多肽处理,并不会明显地影响CLV2-CRN之间的互作强度;进一步的LCI实验发现CLV1不能够与CLV2发生直接的相互作用,但能够与CRN有微弱的相互作用;除此之外,实验人员还发现 CRN自身可以形成同源二聚体,而CLV1CLV2自身不能形成同源二聚体。这些生化和细胞学结果对于新提出的CLV3平行双通路假设提供了直接的证据。

(生物通 小茜)

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Analysis of interactions among the CLAVATA3 receptors reveals a direct interaction between CLAVATA2 and CORYNE in Arabidopsis

Yingfang Zhu 1,2,† , Yuqing Wang 1,2,† , Ruili Li 1,2 , Xiufen Song 1 , Qinli Wang 1,2 , Shanjin Huang 1 , Jing Bo Jin 1 , Chun-Ming Liu 1 and Jinxing Lin 1,*

  1 Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100049, China , and

  2 Gradual School of Chinese Academy of Sciences, Beijing, 100049, China

  * For correspondence (fax +86 10 62836211; e-mail linjx@ibcas.ac.cn).

  † These authors contributed equally to this work.

 

Copyright Journal compilation © 2010 Blackwell Publishing Ltd and the Society for Experimental Biology

KEYWORDS

CORYNE • CLAVATA1 • CLAVATA2 • Arabidopsis mesophyll protoplasts • Nicotiana benthamiana • firefly luciferase complementation imaging assay

ABSTRACT

In Arabidopsis, CORYNE (CRN), a new member of the receptor kinase family, was recently isolated as a key player involved in the CLAVATA3 (CLV3) signaling pathway, thereby playing an important role in regulating the development of shoot and root apical meristems. However, the precise relationships among CLAVATA1 (CLV1), CLAVATA2 (CLV2), and CRN receptors remain unclear. Here, we demonstrate the subcellular localization of CRN and analyze the interactions among CLV1, CLV2, and CRN using firefly luciferase complementation imaging (LCI) assays in both Arabidopsis mesophyll protoplasts and Nicotiana benthamiana leaves. Fluorescence targeting showed that CRN was localized to the plasma membrane. The LCI assays coupled with co-immunoprecipitation assays demonstrated that CLV2 can directly interact with CRN in the absence of CLV3. Additional LCI assays showed that CLV1 did not interact with CLV2, but can interact weakly with CRN. We also found that CLV1 can interact with CLV2–CRN heterodimers, implying that these three proteins may form a complex. Moreover, CRN, rather than CLV1 and CLV2, was able to form homodimers without CLV3 stimulation. Taken together, our results add direct evidence to the newly proposed two-parallel receptor pathways model and therefore provide new insights into the CLV3 signaling pathway.

 

林金星

1992年获北京大学理学博士, 1992-1994年进入中国科学院植物研究所博士后流动站工作。1995年晋升为研究员,1996年获得博士生导师资格。“国家杰出青年基金”获得者,中国科学院“****”入选者。

 

   现在担任全国政协委员、北京市政协委员、北京植物学会理事长,兼任南开大学、中国科大研究生院、北京师范大学、东北师范大学等兼职教授,《Plant Signaling and Behavior》副主编 、《Communicative and Integrative Biology》编委、《科学通报》特约编委、《植物学报》副主编、《植物科学进展》副主编、《电子显微学报》副主编、《植物学通报》编委、中国植物学会植物结构与生殖生物学专业委员会主任等、国家自然科学基金委员会植物学科评审组成员、《中国青年科技奖》评审组成员。

 

代表论文

Nianjun Teng, Biao Jin, Qinli Wang, Huaiqing Hao, Reinhart Ceulemans, Tingyun Kuang, Jinxing Lin*. 2009. No detectable maternal effects of elevated CO2 on Arabidopsis thaliana over 15 generations. PLoS ONE 4(6): e6035. doi:10.1371/journal.pone.0006035.

 

Maozhong Zheng , Martina Beck , Jens Müller , Tong Chen , Xiaohua Wang , Feng Wang , Qinli Wang , Yuqing Wang , František Baluška , David C Logan , Jozef Šamaj , Jinxing lin*. 2009. Actin turnover is

 required for myosin-dependent mitochondrial movements in Arabidopsis root hairs. PLoS ONE 4(6):  

 e5961. doi:10.1371/journal.pone.0005961.

 

 Qiaoling Liu, Bo Chen, Qinli Wang, Xiaoli Shi, Zeyu Xiao, Jinxing Lin, Xiaohong Fang*. 2009. CarbonNanotubes as Molecular Transporters for Walled Plant Cells. Nano Letters 9: 1007–1010. (Highlighted   by Nature China on 04-Mar-2009)

 

Yingfang Zhu, yuqing Wang, Ruili Li, Xiufen Song, Qinli Wang, Shanjin Huang, Jingbo Jin, Chunming Liu, Jinxing Lin*. 2009. Analysis of interactions among the CLAVATA3 receptors reveals a direct interaction between CLAVATA2 and CORYNE in Arabidopsis. The Plant Journal doi: 10.1111/j.1365-313X.2009.04049.x.

 

Peng Liu , Ruili Li, Liang Zhang, Qinli Wang, Karsten Niehaus, Jozef Šamaj, František Baluška, Jinxing Lin*. 2009. Lipid microdomain polarization is required for NADPH oxidase-dependent ROS signaling in Pice meyeri pollen tube tip growth. The Plant Journal  60: 303–313.

 

Tong Chen, Xiaoqin Wu, Yanmei Chen, Xiaojuan Li, Mei Huang, Maozhong Zheng, František Baluška,Jozef Šamaj, Jinxing Lin *. 2009. Combined proteomic and cytological analysis of Ca2+–calmodulin

regulation in Picea meyeri pollen tube growth. Plant Physiology 149: 1111–1126.

 

Yuhua Wang, Tong Chen, Chunyang Zhang, Huaiqing Hao, Peng Liu, Maozhong Zheng, František Baluška,Jozef Šama, Jinxing Lin*. 2009. Nitric oxide modulates the influx of extracellular Ca2+ and actin filamentorganization during cell wall construction in Pinus bungeana pollen tubes. New Phytologist 182: 851–862.

 

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