Perturbations of the ZED1 pseudokinase activate plant immunity

Bastedo, D. Patrick; Khan, Madiha; Martel, Alexandre; Seto, Derek; Kireeva, Inga; Zhang, Jianfeng; Masud, Wardah; Millar, David; Lee, Jee Yeon; Lee, Ahwon; Gong, Yunchen; Santos-Severino, André; Guttman, David S.; Desveaux, Darrell

doi:10.1371/journal.ppat.1007900

Cited by 46 publications

(52 citation statements)

References 92 publications

(186 reference statements)

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“…Although the lack of autoactive mutations for ZAR1 does not allow us to specifically analyze the intramolecular interactions present in active complexes, we demonstrated that associations between the domains must have some flexibility to allow for conformational changes upon activation. Since our interaction assays were carried out in vivo, we also cannot exclude the possibility that other host proteins help to mediate the associations (Bastedo et al, 2019). Our data show that ZAR1 is modulated by subtle conformational changes that are finely controlled, and that ZAR1 displays specificity in its intramolecular interactions despite broad structural similarity with other NLRs.…”

Section: Discussionmentioning

confidence: 92%

“…HopZ1a triggers a strong HR that depends on the RLCK ZED1 (HOPZ-EFFECTOR-TRIGGERED IMMUNITY DEFICIENT 1) and ZAR1 (Lewis et al, 2008(Lewis et al, , 2010(Lewis et al, , 2013Baudin et al, 2017). HopZ1a acetylates ZED1, which is hypothesized to trigger ZAR1 activation, likely through conformational changes (Lewis et al, 2013;Baudin et al, 2017;Bastedo et al, 2019;Wang et al, 2019b). The recognition of AvrAC and HopF2a is more complex and involves other proteins that mediate a connection between the effector and the ZAR1-RLCK complex (Wang et al, 2015b;Seto et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

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Structure–function analysis of ZAR1 immune receptor reveals key molecular interactions for activity

et al. 2019

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Summary NLR (nucleotide‐binding [NB] leucine‐rich repeat [LRR] receptor) proteins are critical for inducing immune responses in response to pathogen proteins, and must be tightly modulated to prevent spurious activation in the absence of a pathogen. The ZAR1 NLR recognizes diverse effector proteins from Pseudomonas syringae, including HopZ1a, and Xanthomonas species. Receptor‐like cytoplasmic kinases (RLCKs) such as ZED1, interact with ZAR1 and provide specificity for different effector proteins, such as HopZ1a. We previously developed a transient expression system in Nicotiana benthamiana that allowed us to demonstrate that ZAR1 function is conserved from the Brassicaceae to the Solanaceae. Here, we combined structural modelling of ZAR1, with molecular and functional assays in our transient system, to show that multiple intramolecular and intermolecular interactions modulate ZAR1 activity. We identified determinants required for the formation of the ZARCC oligomer and its activity. Lastly, we characterized intramolecular interactions between ZAR1 subdomains that participate in keeping ZAR1 immune complexes inactive. This work identifies molecular constraints on immune receptor function and activation.

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Section: Discussionmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Structure–function analysis of ZAR1 immune receptor reveals key molecular interactions for activity

et al. 2019

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“…HopZ1a can acetylate members of a family of PBLs (receptor‐like cytoplasmic kinases) and promote their interaction with ZED1 and ZAR1 to form a ZAR1‐ZED1‐PBL ternary complex. Interactions between ZED1 and PBL kinases are determined by the pseudokinase features of ZED1, and mutants designed to restore ZED1 kinase motifs can bind to PBLs, recruit ZAR1, and trigger ZAR1‐dependent immunity in planta (Bastedo et al , 2019). TaCRK2 may have kinase activity.…”

Section: Discussionmentioning

confidence: 99%

A novel cysteine‐rich receptor‐like kinase gene, TaCRK2, contributes to leaf rust resistance in wheat

Sun

Liu

et al. 2020

Molecular Plant Pathology

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This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. AbstractLeaf rust, caused by Puccinia triticina, is one of the most destructive fungal diseases in wheat production worldwide. The hypersensitive reaction (HR) is an important defence response against P. triticina infection. In this study, the physiological races 165 and 260 of P. triticina were combined with a line derived from the bread wheat cultivar Thatcher with the leaf rust resistance locus Lr26 to form compatible and incompatible combinations, respectively. Based on an RNA-Seq database of the interaction systems, a new wheat cysteine-rich receptor-like kinase gene, TaCRK2, is specifically induced and up-regulated in the incompatible combination. We identified that TaCRK2 was regulated in a Ca 2+ -dependent manner. Knockdown of TaCRK2 by virus-induced gene silencing and RNAi leads to a dramatic increase in HR area and the number of haustorial mother cells at the single infection site. In addition, urediniospores, a P. triticina-specific pathogenic marker in compatible combinations, were observed on leaf surfaces of silenced plants at approximately 15 days after inoculation in the incompatible combination. Moreover, transcription levels of TaPR1, TaPR2, and TaPR5 were obviously reduced in TaCRK2-silenced plants. TaCRK2 overexpression in Nicotiana benthamiana induced strong HR-like cell death. Finally, transient expression of green fluorescent protein fused with TaCRK2 in N. benthamiana indicated that TaCRK2 localizes in the endoplasmic reticulum. Thus, TaCRK2 plays an important role in the resistance to P. triticina infection and has a positive regulation effect on the HR cell death process induced by P. triticina. K E Y W O R D S cysteine-rich receptor-like protein kinase, hypersensitive reaction, Puccinia triticina, VIGS, wheat | 733 GU et al.

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“…SZE1 and SZE2 directly interact with HopZ1a and are proposed to help mediate conformational changes that lead to ZAR1 activation (Liu et al, 2019). In addition, several PBS1‐like kinases (PBLs) have also been shown to be targeted by HopZ1a and participate in ZAR1‐mediated recognition (Bastedo et al, 2019). It would, therefore, be interesting to broaden our diversity screen to these other players in HopZ1a‐induced immunity and investigate how evolution shaped these proteins in relation to ZED1 and ZAR1.…”

Section: Discussionmentioning

confidence: 99%

A natural diversity screen in Arabidopsis thaliana reveals determinants for HopZ1a recognition in the ZAR1‐ZED1 immune complex

Baudin

Martin

Sass

et al. 2020

Plant Cell & Environment

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Pathogen pressure on hosts can lead to the evolution of genes regulating the innate immune response. By characterizing naturally occurring polymorphisms in immune receptors, we can better understand the molecular determinants of pathogen recognition. ZAR1 is an ancient Arabidopsis thaliana NLR (Nucleotide‐binding [NB] Leucine‐rich‐repeat [LRR] Receptor) that recognizes multiple secreted effector proteins from the pathogenic bacteria Pseudomonas syringae and Xanthomonas campestris through its interaction with receptor‐like cytoplasmic kinases (RLCKs). ZAR1 was first identified for its role in recognizing P. syringae effector HopZ1a, through its interaction with the RLCK ZED1. To identify additional determinants of HopZ1a recognition, we performed a computational screen for ecotypes from the 1001 Genomes project that were likely to lack HopZ1a recognition, and tested ~300 ecotypes. We identified ecotypes containing polymorphisms in ZAR1 and ZED1. Using our previously established Nicotiana benthamiana transient assay and Arabidopsis ecotypes, we tested for the effect of naturally occurring polymorphisms on ZAR1 interactions and the immune response. We identified key residues in the NB or LRR domain of ZAR1 that impact the interaction with ZED1. We demonstrate that natural diversity combined with functional assays can help define the molecular determinants and interactions necessary to regulate immune induction in response to pathogens.

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Perturbations of the ZED1 pseudokinase activate plant immunity

Cited by 46 publications

References 92 publications

Structure–function analysis of ZAR1 immune receptor reveals key molecular interactions for activity

Structure–function analysis of ZAR1 immune receptor reveals key molecular interactions for activity

A novel cysteine‐rich receptor‐like kinase gene, TaCRK2, contributes to leaf rust resistance in wheat

A natural diversity screen in Arabidopsis thaliana reveals determinants for HopZ1a recognition in the ZAR1‐ZED1 immune complex

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