Light-triggered and phosphorylation-dependent 14-3-3 association with NONPHOTOTROPIC HYPOCOTYL 3 is required for hypocotyl phototropism

Reuter, Lea; Schmidt, Tatjana; Manishankar, Prabha; Throm, Christian; Keicher, Jutta; Bock, Andrea; Oecking, Claudia

doi:10.1101/2021.04.09.439179

Cited by 2 publications

(4 citation statements)

References 65 publications

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“…Reduction in FLT was not observed when SOBIR1-GFP was co-expressed with SAG101-RFP or plasma membrane-associated RFP-NPH3 S744A (ref. 23 ) (Fig. 3a).…”

Section: Discussionmentioning

confidence: 98%

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The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity

Pruitt

Locci

Wanke

et al. 2021

Nature

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Plants deploy cell-surface and intracellular leucine rich-repeat domain (LRR) immune receptors to detect pathogens 1 . LRR receptor kinases and LRR receptor proteins at the plasma membrane recognize microorganism-derived molecules to elicit pattern-triggered immunity (PTI), whereas nucleotide-binding LRR proteins detect microbial effectors inside cells to confer effector-triggered immunity (ETI). Although PTI and ETI are initiated in different host cell compartments, they rely on the transcriptional activation of similar sets of genes 2 , suggesting pathway convergence upstream of nuclear events. Here we report that PTI triggered by the Arabidopsis LRR receptor protein RLP23 requires signalling-competent dimers of the lipase-like proteins EDS1 and PAD4, and of ADR1 family helper nucleotide-binding LRRs, which are all components of ETI. The cell-surface LRR receptor kinase SOBIR1 links RLP23 with EDS1, PAD4 and ADR1 proteins, suggesting the formation of supramolecular complexes containing PTI receptors and transducers at the inner side of the plasma membrane. We detected similar evolutionary patterns in LRR receptor protein and nucleotide-binding LRR genes across Arabidopsis accessions; overall higher levels of variation in LRR receptor proteins than in LRR receptor kinases are consistent with distinct roles of these two receptor families in plant immunity. We propose that the EDS1-PAD4-ADR1 node is a convergence point for defence signalling cascades, activated by both surface-resident and intracellular LRR receptors, in conferring pathogen immunity.Arabidopsis thaliana (hereafter Arabidopsis) cell-surface LRR receptor kinases (LRR-RKs) and LRR receptor protein (LRR-RP)-SOBIR1 complexes recruit the co-receptor BAK1 and signal through receptor-like cytoplasmic kinases (RLCKs) to elicit PTI 3 . Intracellular coiled-coil (CC)-nucleotide-binding LRR (NLR) or TOLL-INTERLEUKIN 1 RECEP-TOR (TIR)-NLR receptors 4 require ADR1-type and NRG1-type helper NLRs (hNLRs) and the lipase-like EDS1 family proteins EDS1, PAD4 and SAG101 to confer ETI 5,6 . While the defence outputs for PTI and ETI are qualitatively similar 2 , where and how pathways activated in different cell compartments converge remain unclear. Effective plant defence relies on mutual potentiation of PTI and ETI pathways 7,8 , suggesting mechanistic links between these two tiers of the plant immune system. RLCKs PBL30 and PBL31 mediate PTIThe Arabidopsis class VII RLCK (RLCK-VII) BIK1 promotes LRR-RK-mediated PTI but is a negative regulator of LRR-RP-mediated PTI 9 . To identify RLCK-VII members with positive roles in LRR-RP-dependent PTI, we screened an Arabidopsis RLCK-VII transfer DNA mutant library 10 for ethylene production elicited by fungal pg13(At) 11 , oomycete nlp20 and bacterial eMax (which are recognized by RLP42, RLP23 and RLP1, respectively) 3 (Extended Data Fig. 1a). A pbl31 mutant was defective in response to these elicitors compared with wild-type plants (Columbia-0 (Col-0)) (Extended Data Fig. 1a). PBL31 belongs to RLCK-VII subfamily 7, together ...

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“…Reduction in FLT was not observed when SOBIR1-GFP was co-expressed with SAG101-RFP or plasma membrane-associated RFP-NPH3 S744A (ref. 23 ) (Fig. 3a).…”

Section: Discussionmentioning

confidence: 98%

“…N-terminally tagged RFP-NPH3 S744A (in pB7WGR2 (refs. 23,45 )) served as a plasma membrane-associated control; the S744A mutation blocks light-triggered dissociation from the plasma membrane 23 . EDS1-HA and PAD4-HA were expressed from pGWB14 (ref.…”

Section: Fret-flimmentioning

confidence: 99%

The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity

Pruitt

Locci

Wanke

et al. 2021

Nature

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299

263

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“…Lipid overlay assays using PIP strips were performed according to the manufacturer’s instructions (Echelon Biosciences, Salt Lake City, UT, USA) and as described in Reuter et al . (2021).…”

Section: Methodsmentioning

confidence: 99%

“…Protein synthesis was confirmed on Western blot using an haemagglutinin (HA)-specific antibody. Lipid overlay assays using PIP strips were performed according to the manufacturer's instructions (Echelon Biosciences, Salt Lake City, UT, USA) and as described in Reuter et al (2021).…”

Section: Microsomal Fractionationmentioning

confidence: 99%

Arabidopsis ADR1 helper NLR immune receptors localize and function at the plasma membrane in a phospholipid dependent manner

et al. 2021

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Activation of nucleotide-binding leucine-rich repeat receptors (NLRs) results in immunity and a localized cell death. NLR cell death activity requires oligomerization and in some cases plasma membrane (PM) localization. The exact mechanisms underlying PM localization of NLRs lacking predicted transmembrane domains or recognizable lipidation motifs remain elusive.We used confocal microscopy, genetically encoded molecular tools and protein-lipid overlay assays to determine whether PM localization of members of the Arabidopsis HeLo-/ RPW8-like domain 'helper' NLR (RNL) family is mediated by the interaction with negatively charged phospholipids of the PM.Our results show that PM localization and stability of some RNLs and one CC-type NLR (CNL) depend on the direct interaction with PM phospholipids. Depletion of phosphatidylinositol-4-phosphate from the PM led to a mis-localization of the analysed NLRs and consequently inhibited their cell death activity. We further demonstrate homo-and hetero-association of members of the RNL family. Our results provide new insights into the molecular mechanism of NLR localization and defines an important role of phospholipids for CNL and RNL PM localization and consequently, for their function.We propose that RNLs interact with anionic PM phospholipids and that RNL-mediated cell death and immune responses happen at the PM.

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Light-triggered and phosphorylation-dependent 14-3-3 association with NONPHOTOTROPIC HYPOCOTYL 3 is required for hypocotyl phototropism

Cited by 2 publications

References 65 publications

The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity

The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity

Arabidopsis ADR1 helper NLR immune receptors localize and function at the plasma membrane in a phospholipid dependent manner

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