Identification and receptor mechanism of TIR-catalyzed small molecules in plant immunity

Science, Journal Year: 2022, Volume and Issue: 377(6605)

Published: July 7, 2022

Plant nucleotide-binding leucine-rich repeat-containing (NLR) receptors with an N-terminal Toll/interleukin-1 receptor (TIR) domain sense pathogen effectors to enable TIR-encoded nicotinamide adenine dinucleotide hydrolase (NADase) activity for immune signaling. TIR-NLR signaling requires the helper NLRs N requirement gene 1 (NRG1), Activated Disease Resistance (ADR1), and Enhanced Susceptibility (EDS1), which forms a heterodimer each of its paralogs Phytoalexin Deficient 4 (PAD4) Senescence-Associated Gene 101 (SAG101). Here, we show that TIR-containing proteins catalyze production 2'-(5''-phosphoribosyl)-5'-adenosine monophosphate (pRib-AMP) diphosphate (pRib-ADP) in vitro planta. Biochemical structural data demonstrate EDS1-PAD4 is complex pRib-AMP pRib-ADP, allosterically promote interaction ADR1-L1 but not NRG1A. Our study identifies TIR-catalyzed pRib-ADP as missing link TIR through likely second messengers plant immunity.

Language: Английский

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TIR-catalyzed ADP-ribosylation reactions produce signaling molecules for plant immunity

Science, Journal Year: 2022, Volume and Issue: 377(6605)

Published: July 7, 2022

Plant pathogen-activated immune signaling by nucleotide-binding leucine-rich repeat (NLR) receptors with an N-terminal Toll/interleukin-1 receptor (TIR) domain converges on Enhanced Disease Susceptibility 1 (EDS1) and its direct partners, Phytoalexin Deficient 4 (PAD4) or Senescence-Associated Gene 101 (SAG101). TIR-encoded nicotinamide adenine dinucleotide hydrolase (NADase) produces molecules to promote exclusive EDS1-PAD4 EDS1-SAG101 interactions helper NLR subclasses. In this work, we show that TIR-containing proteins catalyze adenosine diphosphate (ADP)-ribosylation of triphosphate (ATP) ADP ribose (ADPR) through ADPR polymerase-like NADase activity, forming ADP-ribosylated ATP (ADPr-ATP) ADPr-ADPR (di-ADPR), respectively. Specific binding ADPr-ATP di-ADPR allosterically promotes interaction N requirement gene 1A (NRG1A) in vitro planta. Our data reveal enzymatic activity TIRs enables specific activation the EDS1-SAG101-NRG1 immunity branch.

Language: Английский

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NOD-like receptor-mediated plant immunity: from structure to cell death

Nature reviews. Immunology, Journal Year: 2020, Volume and Issue: 21(5), P. 305 - 318

Published: Dec. 8, 2020

Language: Английский

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Short prokaryotic Argonaute systems trigger cell death upon detection of invading DNA

Cell, Journal Year: 2022, Volume and Issue: 185(9), P. 1471 - 1486.e19

Published: April 1, 2022

Argonaute proteins use single-stranded RNA or DNA guides to target complementary nucleic acids. This allows eukaryotic mediate interference and long prokaryotic interfere with invading The function mechanisms of the phylogenetically distinct short remain poorly understood. We demonstrate that associated TIR-APAZ (SPARTA) form heterodimeric complexes. Upon guide RNA-mediated binding, four SPARTA heterodimers oligomers in which TIR domain-mediated NAD(P)ase activity is unleashed. When expressed Escherichia coli, activated presence highly transcribed multicopy plasmid DNA, causes cell death through NAD(P)+ depletion. results removal plasmid-invaded cells from bacterial cultures. Furthermore, we show can be repurposed for programmable detection sequences. In conclusion, our work identifies as a immune system reduces viability upon RNA-guided DNA.

Language: Английский

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TIR domains of plant immune receptors are 2′,3′-cAMP/cGMP synthetases mediating cell death

Cell, Journal Year: 2022, Volume and Issue: 185(13), P. 2370 - 2386.e18

Published: May 20, 2022

Language: Английский

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Pathogen effector recognition-dependent association of NRG1 with EDS1 and SAG101 in TNL receptor immunity

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: June 7, 2021

Plants utilise intracellular nucleotide-binding, leucine-rich repeat (NLR) immune receptors to detect pathogen effectors and activate local systemic defence. NRG1 ADR1 "helper" NLRs (RNLs) cooperate with enhanced disease susceptibility 1 (EDS1), senescence-associated gene 101 (SAG101) phytoalexin-deficient 4 (PAD4) lipase-like proteins mediate signalling from TIR domain NLR (TNLs). The mechanism of RNL/EDS1 family protein cooperation is not understood. Here, we present genetic molecular evidence for exclusive EDS1/SAG101/NRG1 EDS1/PAD4/ADR1 co-functions in TNL immunity. Using immunoprecipitation mass spectrometry, show effector recognition-dependent interaction EDS1 SAG101, but PAD4. An EDS1-SAG101 complex interacts NRG1, EDS1-PAD4 ADR1, an immune-activated state. requires intact nucleotide-binding P-loop motif, a functional EP its partner induced association Thus, two distinct modules (NRG1/EDS1/SAG101 ADR1/EDS1/PAD4) receptor defence signalling.

Language: Английский

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RefPlantNLR is a comprehensive collection of experimentally validated plant disease resistance proteins from the NLR family

PLoS Biology, Journal Year: 2021, Volume and Issue: 19(10), P. e3001124 - e3001124

Published: Oct. 20, 2021

Reference datasets are critical in computational biology. They help define canonical biological features and essential for benchmarking studies. Here, we describe a comprehensive reference dataset of experimentally validated plant nucleotide-binding leucine-rich repeat (NLR) immune receptors. RefPlantNLR consists 481 NLRs from 31 genera belonging to 11 orders flowering plants. This has several applications. We used determine the functionally benchmark 5 NLR annotation tools. revealed that although tools tend retrieve majority NLRs, they frequently produce domain architectures inconsistent with annotation. Guided by this analysis, developed new pipeline, NLRtracker, which extracts annotates protein or transcript files based on core found dataset. The should also prove useful guiding comparative analyses across wide spectrum diversity identifying understudied taxa. hope resource will contribute moving field beyond uniform view structure function.

Language: Английский

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Ca ²⁺ signals in plant immunity

The EMBO Journal, Journal Year: 2022, Volume and Issue: 41(12)

Published: May 13, 2022

Review13 May 2022Open Access Ca2+ signals in plant immunity Philipp Köster orcid.org/0000-0002-1359-822X Institute of Plant and Microbial Biology Zürich-Basel Science Center, University Zürich, Switzerland Contribution: Visualization, Writing - original draft, review & editing Search for more papers by this author Thomas A DeFalco orcid.org/0000-0003-2897-1485 Cyril Zipfel Corresponding Author [email protected] orcid.org/0000-0003-4935-8583 The Sainsbury Laboratory, East Anglia, Norwich, UK Funding acquisition, Information Köster1, DeFalco1 *,1,2 1Institute 2The *Corresponding author. Tel: +41 044 63 48222; E-mail: EMBO Journal (2022)41:e110741https://doi.org/10.15252/embj.2022110741 PDFDownload PDF article text main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures Info Abstract Calcium ions function as a key second messenger ion eukaryotes. Spatially temporally defined cytoplasmic are shaped through the concerted activity channels, exchangers, pumps response diverse stimuli; these then decoded Ca2+-binding sensor proteins. In plants, signaling is central both pattern- effector-triggered immunity, with generation characteristic elevations potential pathogens being common both. However, despite their importance, long history scientific interest, transport proteins that shape integration remain poorly characterized. Here, we discuss recent work has shed light on deepened mysteries immunity. immune system All eukaryotes use systems protect themselves against pathogens. consists two characterized perception layers: one utilizes cell-surface pattern recognition receptors (PRRs) perceive extracellular immunogenic patterns, another relies intracellular nucleotide-binding leucine-rich repeat (NLR) recognize pathogenic effectors inside cell (Jones Dangl, 2006). first layer system, apoplastic elicitors such pathogen-, microbe-, damage-, or herbivore-associated molecular patterns (PAMPs, MAMPs, DAMPs, HAMPs, respectively) immune-modulating peptide phytocytokines recognized PRRs, which leads defense responses termed pattern-triggered (PTI) (Boller Felix, 2009; Yu et al, 2017; Zipfel, 2021). PRRs described date receptor kinases (RKs) (RPs) (Boutrot Albert 2020). RKs domain structure reminiscent metazoan tyrosine (RTKs) (DeFalco 2021); namely, ligand-binding (ECD), single-span transmembrane helix (TM) cytosolic protein kinase (Jamieson 2018), while RPs lack instead form functional bipartite adapter (Liebrand 2013; 2015; Postma 2016). Because architecture, plasma membrane (PM)-localized (or complexes) allow ligand binding be communicated across into events. nature varies, including proteins, lipids, carbohydrates, can derived from either pathogen herbivore (e.g., PAMPs, HAMPs) host plant, case macromolecules released upon damage (DAMPs) secreted (Gust 2017). PRR ECDs variety subdomains, (LRR), epidermal growth factor-like (EGF), lectin, lysin motif (LysM) domains best-studied to-date LRR-RKs FLAGELLIN-SENSING 2 (FLS2) EF-TU RECEPTOR (EFR), bacterial PAMPs flg22 elf18, respectively (Gómez-Gómez Boller, 2000; Both FLS2 EFR stable ligand-dependent complexes LRR-RK co-receptors SOMATIC EMBRYOGENESIS KINASE (SERK) family, BRASSINOSTEROID-INSENSITIVE 1-ASSOCIATED 1 (BAK1, also called SERK3) (Chinchilla 2007; Heese Roux 2011). Complex formation between phosphorylation events within activation receptor-like (RLCKs), directly phosphorylate regulate target order activate PTI (Liang Zhou, 2018; 2021) (Fig 1A). Figure 1. ETI induce residing at PM. They originating microbes, viruses, herbivores, parasitic damaged cells. turn, RLCKs activated downstream release few minutes after facet. Microbes introduce effector cells disturb overcome responses. Cytoplasmic NLRs sense presence ETI. To end, autoinhibition released, ADP changed ATP oligomerization occurs, leading finally (A). significant increase been reported occur Arabidopsis leaves starting 1.5 h peaking about infection avirulent bacteria (B). Schematic signatures plants induced Grant al (2000) RK: kinase; co-RK: coreceptor RP: protein; RLCK: like NLR: receptor; CC: coiled-coil; TIR: toll/interleukin-related; CNLs: CC-NLRs; TNLs: TIR-NLRs; RNLs: RPW8-NRLs; NBS: nucleotide site; LRR: repeats; PTI: immunity; ETI: c[Ca2+]: free concentration. Download figure PowerPoint Pathogens cytoplasm promote pathogenicity, often disturbing counteract this, rely NLR-type and/or activity, (ETI). Interestingly, share architecture those animals, featuring conserved (NBD) LRR domain, variable accessory N C termini (DeYoung Innes, 2006; Jones 2016; Baggs van Wersch categorized based N-terminal domains: coiled-coil (CC)-NLRs (CNLs), toll/interleukin-related (TIR)-NLRs (TNLs), RPW8-NLRs (RNLs). Of NLRs, CNLs TNLs sensors RNLs helpers (Baggs Wu Jubic 2019; Feehan present an inactive state, likely autoinhibitory, adenosine diphosphate (ADP) bound NBD (Williams 2011; Bernoux Upon activation, exchanged triphosphate (ATP) NLR via large multimeric structures (Danot 2009). similar mechanism hypothesized but only recently corroborated structural data discussed detail below. have traditionally viewed independent pathways; however, least some components shared layers (Thomma Activation triggers numerous overlapping events, fluxes, production reactive oxygen species (ROS), mitogen-activated (MAPK) cascades, transcriptional reprogramming, phytohormone biosynthesis (Cui Zhou Zhang, 2020; generally accompanied programmed death hypersensitive (HR) site 2006), although HR-like forms (Wang Recent further demonstrated linked levels (Ngou 2021; Pruitt Tian Yuan exact mechanisms governing linkage pathways remains elucidated fully. As changes intracelluar well documented thought (Seybold 2014; Moeder 2019). universal (Clapham, 2007). Owing its cytotoxicity, must maintained low (~10−8 10−7 M) living cells, thus sequestered stores (in primarily vacuole endoplasmatic reticulum, vesicular compartments, chloroplasts mitochondria) apoplast active transport, generating enormous electrochemical gradients membranes Edel Costa 2018). Ca2+-permeable channels therefore generate rapid, transient increases concentrations, turn interpreted suite cellular processes 2010). summarized three steps: encoding (via stimulus-triggered fluxes), decoding proteins), regulation processes). involved all aspects life, regulation, development, abiotic stress responses, reproduction (Kudla establishment beneficial plant-microbe interactions (Tian review, focus how encoded PM during signaling. influx oxidative burst (Doke, 1983, 1985; Apostol 1989; Keppler 1989) were among elicitor treatment (Atkinson 1996; Levine Zimmermann 1997; Lecourieux 2002). ROS was eventually attributed PM-localized NADPH oxidases RESPIRATORY BURST OXIDASE HOMOLOGUE (RBOH) family (Torres 2002); model thaliana (hereafter, Arabidopsis), single member, RBOHD, responsible (Nühse Zhang contrast, channel(s) remained comparably elusive many years 2014). Cytosolic evoked various measured culture using radioisotopes, Ca2+-sensitive dyes, electrophysiological approaches Gelli 1997). development genetically indicators (GECIs) greatly expanded possibilities real-time, kinetic analysis fluxes intact tissues treatment. GECI deployed aequorin (AEQ) Aequoria victoria (Knight 1991), holo-enzyme cofactor coelenterazine emits Ca2+-binding. When challenged virulent strains bacterium Pseudomonas syringae, expressing AEQ showed signal peak ~10 min. second, stronger, persistent seen 1.5–2 avirulent, ETI-activating P. syringae (Grant Kang 2010; Hung kinetics early elevation triggered (Blume 2002) biphasic ETI-inducing suggested may distinct 1B). Subsequent analyses AEQ-expressing shown elicitors, phytocytokines, sufficient elicit rapid (Ranf 2008, Vadassery Krol Such requires components, RLCK-VII/ AVRPPHB SUSCEPTIBLE (PBS1)-LIKE (PBL) members BOTRYTIS-INDUCED (BIK1) PBL1 (Li Ranf Monaghan 2015). More recently, deployment fluorescent GECIs allowed elicitor-induced level. include ratiometric yellow cameleons) intensiometric GCaMPs GECOs) (Grenzi 2021b; Waadt Flourescent utilized show oscillatory single-cell level (Thor Peiter, Keinath 2015) roots application laser ablation-induced lead transients (Keinath Marhavý Ca2+—tightly messengers There extensive interplay (Gilroy 2016); initial PTI-related mildly reduced oxidase inhibitor DPI catalase, there no effect longer-term, 2000). Similarly, rbohd mutants slight, quantitative defect elicitor-triggered when seedlings severely attenuated channel blockers Elicitor RBOHD BIK1 (Kadota Li 2014), suggesting complex relationship wherein, perception, PRR-mediated primes subsequent 2). not activates EF-hand indirectly Ca2+-regulated kinase-mediated (Ogasawara 2008; Dubiella 2013). CALCIUM DEPENDENT PROTEIN 5 (CPK5) sites (Dubiella Kadota While residues strictly required PTI-induced bursts 2007), individual contribution other impact certain uncovered. 2. tightly interconnected PTI, signals. NADPH-oxidase EF-hands, terminus (indicated grey arrows targeting p-sites). addition, thereby black Reactive perceived cysteine pairs RK HPCA1/CARD1. This H2O2 Arabidopsis, pathway HPCA1 known. AEQ-based screen impaired H2O2-induced identified LRR-RK, HYDROGEN PEROXIDE INDUCED INCREASE (HPCA1), putative (Wu 2020a). independently CANNOT RESPOND TO DMBQ (CARD1), loss quinone compound 2,6-dimethoxy-1,4-benzoquinone (DMBQ), regulates HPCA1/CARD1-dependent (Laohavisit regulated HPCA1/CARD1, role regulating sensor(s) unclear. AEQ-measured calcium cngc2 cngc4 2019), perception. Shaping efflux generated coordinated action transporters involve (Spalding Harper, Resentini possess major families mediate out cytosol: Ca2+/H+ exchangers (CAXs), autoinhibited Ca2+-ATPases (ACAs) ER (Geisler Shigaki Hirschi, García Bossi ACA relieved Ca2+/CaM-binding, allows feedback ACA8 homolog ACA10 interactors FLS2, aca8 aca10 displayed defects flg22-induced compromised resistance (Frei dit Frey 2012), disturbed stomatal closure PAMP (Yang 2017), shapes PTI. Two tonoplast-localized ACAs, ACA4 ACA11, implicated aca4 aca11 display autoimmune phenotypes spontaneous (Boursiac Although wildtype total content 2010), revealed basal elevated (Hilleary Elicitor-induced peaks 3), rescued mis-localization ACAs tonoplast 2020), indicating critical maintain homeostasis modulate 3. Disturbance machinery impairs (CAX) Ca2+-ATPase reside establish concentrations termination export vacuolar lumen mutants, consequently phenotype lines, slower onset signal, higher concentration retarded reduction Hilleary (2020) (C). Plasma membrane-localized Extensive require PM-localized, Gd3+ La3+ abolishes 2002; Kwaaitaal Maintz studies clearly implicate signaling, hidden. started decipher defense-related roles several classes begun Below, immunity-related candidates phylogenetic groups rather than following chronological identification strict PTI/ETI dichotomy. CNGCs—from strong One tetrameric cyclic nucleotide-gated (CNGCs) (Köhler Neuhaus, 1998). CNGCs comprise gene 20 Arabidopsis) (Mäser 2001) named topology organization, mammalian (CNG) hyperpolarization-activated nucleotide-modulated (HCN) (Kaupp Seifert, Matulef Zagotta, 2003). Individual six helices termini, (CNBD) located CNGC (Kaplan previous reports indicated CNBDs bind nucleotides (Baxter 2008), cAMP cGMP (Leng Gao 2014, Meena it unclear whether bona fide agonists planta. Furthermore, existence guanylate adenylate cyclases (GCs ACs) proteomes still under debate will here. Indeed, suggest multiple RKs, GC (Qi Turek Irving, 2021), determined vitro activities GCs position argues physiological relevance (Ashton, Bojar Nevertheless, over past decades Ca2+-permeable, non-selective cation (Jarratt-Barnham calmodulin (CaM), CaM-binding (CaMBDs) found examined (Arazi 1999; Köhler Hua 2003; Fischer 2013, 2016a) isoforms 2016a). Ca2+/CaM 2016b) Ca2+-independent IQ CaMBD C-terminal end essential 2016a; Pan additional Ca2+-dependent CaMBDs providing negative (feedback) divided four subfamilies phylogeny, group IV IVa IVb 2001). members, CNGC2 CNGC4, isolated defense, (dnd) lesion mimic (hlm) dnd1 dnd2/hlm1 (null (Clough Balagué Jurkowski 2004). dnd initially defective induction HR, able carry (Yu These phenotypic defects, dwarf morphology, delayed flowering, salicylic acid (SA), death, dis-regulated auxin Chan 2004; Chin Chakraborty keeping immune-related dnd1/cngc2 mediator molecule nitric oxide (NO) compared WT lipopolysaccharide (LPS) (Ali same study used pharmacological inhibitors CaM, NO synthase (NOS)-type process. Given mammalian-type NOS enzymes land (Santolini 2017) myriad functions CaM results however cautiously. reporter lines full (Ma 2012). convergence (Couto Bjornson specificity achieved. virus-induced silencing (VIGS) tomato flg22, positively (Saand Recently, loss-of-function each forward genetic exhibited syringae. Remarkably, dependent high media, indistinguishable plants. bik1, do conditional Detailed characterization heterologously expressed Xenopus laevis oocytes subunits inactive, CNGC2-CNGC4 heteromers produce currents wherein together (Chin inhibited CaM; experiments CNGC4 partially 2019) 4A). highlights subject, phosphorylation, and, potentially, 4. fulfil homo- heterotertramers homotetramers heterotetramers cytosol. Ca2+-bound Calmodulin (CAM) inhibits loop. initiation phosphorylates CAM-mediated inhibition rice, RLCK185 OsCNGC9. If OsCNGC9 containing tetramer homomeric heteromeric known resolved pathways. CNGC19 CNGC20 PM, phosphorylated BAK1, initiates degradation channels. bak1/bkk1 coRK accumulation CNGC19/CNGC20 influx, ultimately causing death. media (Chan 2013),

Language: Английский

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Long-read genome sequencing of bread wheat facilitates disease resistance gene cloning

Nature Genetics, Journal Year: 2022, Volume and Issue: 54(3), P. 227 - 231

Published: March 1, 2022

Abstract The cloning of agronomically important genes from large, complex crop genomes remains challenging. Here we generate a 14.7 gigabase chromosome-scale assembly the South African bread wheat ( Triticum aestivum ) cultivar Kariega by combining high-fidelity long reads, optical mapping and chromosome conformation capture. resulting is an order magnitude more contiguous than previous assemblies. shows durable resistance to devastating fungal stripe rust disease 1 . We identified race-specific gene Yr27 , which encodes intracellular immune receptor, be major contributor this resistance. allelic leaf Lr13 ; proteins show 97% sequence identity 2,3 Our results demonstrate feasibility generating assemblies clone genes, exemplify that highly similar alleles single-copy can confer different pathogens, might provide basis for engineering with multiple recognition specificities in future.

Language: Английский

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Recent Advances in Effector-Triggered Immunity in Plants: New Pieces in the Puzzle Create a Different Paradigm

International Journal of Molecular Sciences, Journal Year: 2021, Volume and Issue: 22(9), P. 4709 - 4709

Published: April 29, 2021

Plants rely on multiple immune systems to protect themselves from pathogens. When pattern-triggered immunity (PTI)-the first layer of the response-is no longer effective as a result pathogenic effectors, effector-triggered (ETI) often provides resistance. In ETI, host plants directly or indirectly perceive pathogen effectors via resistance proteins and launch more robust rapid defense response. Resistance are typically found in form nucleotide-binding leucine-rich-repeat-containing receptors (NLRs). Upon effector recognition, an NLR undergoes structural change associates with other NLRs. The dimerization oligomerization NLRs signals downstream components, activates "helper" NLRs, culminates ETI Originally, PTI was thought contribute little ETI. However, most recent studies revealed crosstalk cooperation between PTI. Here, we summarize advancements our understanding response its well how these components cooperate innate signaling pathways. Based up-to-date accumulated knowledge, this review current perspective potential engineering strategies for crop protection.

Language: Английский

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