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RNAi-based screen reveals concerted functions of MEKK2 and CRCK3 in plant cell death regulation.

Authors
  • Yang, Yong1
  • Liu, Jun2
  • Yin, Chuanchun3
  • Vespoli, Luciano de Souza3
  • Ge, Dongdong3
  • Huang, Yanyan4
  • Feng, Baomin2
  • Xu, Guangyuan2
  • Manhães, Ana Marcia E de A3
  • Dou, Shijuan2
  • Criswell, Cameron2
  • Shan, Libo5
  • Wang, Xiaofeng6
  • He, Ping7
  • 1 State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University CITY: Yangling STATE: Shaanxi China [CN]. , (China)
  • 2 Department of Biochemistry & Biophysics, Institute for Plant Genomics & Biotechnology, Texas A&M University CITY: College Station STATE: Texas United States Of America [US]. , (United States)
  • 3 Department of Plant Pathology & Microbiology, Institute for Plant Genomics & Biotechnology, Texas A&M University CITY: College Station STATE: Texas United States Of America [US]. , (United States)
  • 4 Texas A&M University CITY: College Station STATE: Texas United States Of America [US]. , (United States)
  • 5 Texas A&M University CITY: College Station STATE: Texas POSTAL_CODE: 77843 United States Of America [US]. , (United States)
  • 6 State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University CITY: yangling STATE: shaanxi POSTAL_CODE: 712100 China [CN]. , (China)
  • 7 Texas A&M University CITY: College Station STATE: Texas POSTAL_CODE: 77843 United States Of America [US] [email protected] , (United States)
Type
Published Article
Journal
Plant physiology
Publication Date
Mar 12, 2020
Identifiers
DOI: 10.1104/pp.19.01555
PMID: 32165446
Source
Medline
Language
English
License
Unknown

Abstract

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 RNAi-based screen using a virus-induced gene silencing (VIGS) 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 MEKK1, MKK1/2, and MPK4, which depends on a nucleotide-binding site leucine-rich repeat (NLR) protein SUMM2. Silencing of MEKK1 by VIGS resembles the mekk1 mutant with autoimmunity and defense activation. The RNAi-based screen towards Arabidopsis T-DNA insertion lines identified SUMM2, MAPK kinase kinase 2 (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 upregulated 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. {copyright, serif} 2020 American Society of Plant Biologists. All rights reserved.

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