当前位置: X-MOL 学术Plant Physiol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
RNA Interference-Based Screen Reveals Concerted Functions of MEKK2 and CRCK3 in Plant Cell Death Regulation.
Plant Physiology ( IF 7.4 ) Pub Date : 2020-03-12 , DOI: 10.1104/pp.19.01555
Yong Yang 1, 2 , Jun Liu 3 , Chuanchun Yin 2 , Luciano de Souza Vespoli 2 , Dongdong Ge 2 , Yanyan Huang 3 , Baomin Feng 3 , Guangyuan Xu 3 , Ana Marcia E de A Manhães 2 , Shijuan Dou 3, 4 , Cameron Criswell 3 , Libo Shan 2 , Xiaofeng Wang 1 , Ping He 5
Affiliation  

A wide variety of intrinsic and extrinsic cues lead to cell death with unclear mechanisms. The infertility of some death mutants often hurdles the classical suppressor screens for death regulators. We have developed a transient RNA interference (RNAi)-based screen using a virus-induced gene silencing approach to understand diverse cell death pathways in Arabidopsis (Arabidopsis thaliana). One death pathway is due to the depletion of a MAP kinase (MAPK) cascade, consisting of MAPK kinase kinase 1 (MEKK1), MKK1/2, and MPK4, which depends on a nucleotide-binding site Leu-rich repeat (NLR) protein SUMM2. Silencing of MEKK1 by virus-induced gene silencing resembles the mekk1 mutant with autoimmunity and defense activation. The RNAi-based screen toward Arabidopsis T-DNA insertion lines identified SUMM2, MEKK2, and Calmodulin-binding receptor-like cytoplasmic kinase 3 (CRCK3) to be vital regulators of RNAi MEKK1-induced cell death, consistent with the reports of their requirement in the mekk1-mkk1/2-mpk4 death pathway. Similar with MEKK2, overexpression of CRCK3 caused dosage- and SUMM2-dependent cell death, and the transcripts of CRCK3 were up-regulated in mekk1, mkk1/2, and mpk4 MEKK2-induced cell death depends on CRCK3. Interestingly, CRCK3-induced cell death also depends on MEKK2, consistent with the biochemical data that MEKK2 complexes with CRCK3. Furthermore, the kinase activity of CRCK3 is essential, whereas the kinase activity of MEKK2 is dispensable, for triggering cell death. Our studies suggest that MEKK2 and CRCK3 exert concerted functions in the control of NLR SUMM2 activation and MEKK2 may play a structural role, rather than function as a kinase, in regulating CRCK3 protein stability.

中文翻译:

基于 RNA 干扰的筛选揭示了 MEKK2 和 CRCK3 在植物细胞死亡调节中的协同功能。

多种内在和外在线索导致细胞死亡,但机制尚不清楚。一些死亡突变体的不育性常常阻碍死亡调节因子的经典抑制筛选。我们开发了一种基于瞬时 RNA 干扰 (RNAi) 的筛选方法,使用病毒诱导的基因沉默方法来了解拟南芥 (Arabidopsis thaliana) 中不同的细胞死亡途径。一种死亡途径是由于 MAP 激酶 (MAPK) 级联的耗尽所致,该级联由 MAPK 激酶激酶 1 (MEKK1)、MKK1/2 和 MPK4 组成,依赖于核苷酸结合位点富含亮氨酸的重复序列 (NLR) 蛋白总和2。病毒诱导的基因沉默对 MEKK1 的沉默类似于具有自身免疫和防御激活的 mekk1 突变体。基于 RNAi 的拟南芥 T-DNA 插入系筛选发现 SUMM2、MEKK2 和钙调蛋白结合受体样细胞质激酶 3 (CRCK3) 是 RNAi MEKK1 诱导的细胞死亡的重要调节因子,这与它们在mekk1-mkk1/2-mpk4 死亡途径。与MEKK2类似,CRCK3的过表达引起剂量和SUMM2依赖性细胞死亡,并且CRCK3的转录本在mekk1、mkk1/2和mpk4中上调,MEKK2诱导的细胞死亡依赖于CRCK3。有趣的是,CRCK3诱导的细胞死亡也依赖于MEKK2,这与MEKK2与CRCK3复合的生化数据一致。此外,对于触发细胞死亡,CRCK3的激酶活性是必需的,而MEKK2的激酶活性是可有可无的。我们的研究表明,MEKK2 和 CRCK3 在控制 NLR SUMM2 激活方面发挥协同作用,并且 MEKK2 在调节 CRCK3 蛋白稳定性方面可能发挥结构作用,而不是作为激酶发挥作用。
更新日期:2020-05-01
down
wechat
bug