Plant and prokaryotic TIR domains generate distinct cyclic ADPR NADase products

Science Advances, Journal Year: 2023, Volume and Issue: 9(11)

Published: March 17, 2023

Toll/interleukin-1 receptor (TIR) domain proteins function in cell death and immunity. In plants bacteria, TIR domains are often enzymes that produce isomers of cyclic adenosine 5′-diphosphate–ribose (cADPR) as putative immune signaling molecules. The identity functional conservation cADPR isomer signals is unclear. A previous report found a plant could cross-activate the prokaryotic Thoeris TIR–immune system, suggesting TIR-immune signals. Here, we generate autoactive TIRs test converse hypothesis: Do also immunity? Using planta vitro assays, find overlapping sets further clarify how activate system via producing 3′cADPR. This study demonstrates requirements for systems distinct across kingdoms diversity small-molecule products.

Language: Английский

---

NLR signaling in plants: from resistosomes to second messengers

Trends in Biochemical Sciences, Journal Year: 2023, Volume and Issue: 48(9), P. 776 - 787

Published: June 30, 2023

Pathogen effector-induced assembly of resistosomes has been established as an important event for nucleotide binding and leucine-rich repeat-containing receptor (NLR) signaling in plants.The pentameric coiled-coil domain-containing NLR (CNL) act Ca2+-permeable channels, whereas the tetrameric Toll-interleukin 1-like (TIR) (TNL) are NADase holoenzymes.TNL catalyze production nucleotide-derived second messengers to activate downstream helper NLRs activated disease resistance 1 (ADR1) N requirement gene (NRG1) CNL class. Thus, CNLs TNLs converge on Ca2+ signals trigger plant immunity.NLR cross-talks with pattern-triggered immunity (PTI) pathways.NLR pathways plants negatively regulated by both hosts pathogens. Nucleotide receptors (NLRs) have a critical role through direct or indirect recognition pathogen effectors. Recent studies demonstrated that such induces formation large protein complexes called mediate immune signaling. Some influx acting others function active NADases messengers. In this review we summarize these resistosome-mediated derivatives. We also discuss events regulation resistosome Plants rely multiple detect invading microbial pathogens mount responses [1.Ngou B.P.M. et al.Thirty years resistance: zig-zag system.Plant Cell. 2022; 34: 1447-1478Crossref PubMed Scopus (45) Google Scholar,2.Zhou J.M. Zhang Y. Plant immunity: danger perception signaling.Cell. 2020; 181: 978-989Abstract Full Text PDF (328) Scholar]. One subfamily pattern-recognition (PRRs) (see Glossary) at cell surface Scholar, 2.Zhou 3.DeFalco T.A. Zipfel C. Molecular mechanisms early signaling.Mol. 2021; 81: 3449-3467Abstract (0) PRRs recognize pathogen-associated molecular patterns (PAMPs) host-derived damage-associated (DAMPs), leading (PTI). PTI constitutes first line inducible defense against can breach layer secreting effector proteins into cells dampen PTI. To counteract virulence activity effectors, evolved receptors: intracellular NLRs. specifically proteins, inducing effector-triggered (ETI) confer race-specific site entry Scholar,4.Jones J.D.G. al.Intracellular innate surveillance devices animals.Science. 2016; 354aaf6395Crossref different structures subcellular localizations but conserved responses, including influx, bursts reactive oxygen species (ROS), phytocytokines phytohormones, transcriptional reprogramming Probably reason, ETI tightly connected [5.Yuan M. al.Pattern-recognition required NLR-mediated immunity.Nature. 592: 105-109Crossref (351) Scholar,6.Ngou al.Mutual potentiation cell-surface receptors.Nature. 110-115Crossref (329) However, differ timing, amplitude, duration defense, which could be determining their physiological outcomes. addition includes hypersensitive response (HR), form rapid localized programmed death infection [7.Jones J.D. Dangl J.L. The system.Nature. 2006; 444: 323-329Crossref (8693) largest hundreds distinct members [4.Jones two domains: central nucleotide-binding oligomerization domain (NOD), C-terminal repeat (LRR) domain. A variable (CC) is attached terminus, resulting TIR-NLR CC-NLR (CNL), respectively [8.Maruta N. al.Structural basis activation signalling plants.Immunogenetics. 74: 5-26Crossref (25) Scholar,9.Hu Z. Chai J. Assembly architecture inflammasomes.Annu. Rev. Biophys. 52: 8.1-8.22Google pathogen-sensing NLRs, there some (h) translate from [10.Feehan al.Plant get little help friends.Curr. Opin. Biol. 56: 99-108Crossref (18) Scholar,11.Jubic L.M. al.Help wanted: responses.Curr. 2019; 50: 82-94Crossref (139) Examples hNLRs include Resistance Powdery Mildew 8 (RPW8) family [12.Wu al.Differential TNL-mediated redundant CNLs.New Phytol. 222: 938-953Crossref (118) 13.Castel B. al.Diverse defence via RPW8-NLR NRG1.New 966-980Crossref (144) 14.Qi T. al.NRG1 functions EDS1 regulate TIR-NLR-mediated Nicotiana benthamiana.Proc. Natl. Acad. Sci. U. S. A. 2018; 115: E10979-E10987Crossref (121) 15.Collier S.M. al.Cell mediated N-terminal domains unique highly class NB-LRR protein.Mol. Microbe Interact. 2011; 24: 918-931Crossref (230) Scholar] (Figure 1) (NRCs) [16.Adachi H. Kamoun networks plants.Essays Biochem. 66: 541-549Crossref (4) Scholar,17.Wu C.H. al.NLR network mediates diverse pathogens.Proc. 2017; 114: 8113-8118Crossref (211) More recent signaling, many sensor Solanaceae, depends NRCs upon [18.Ahn H.K. al.Effector-dependent NRC Rpi-amr3 Rpi-amr1.EMBO 2023; 42e111484Crossref (5) Scholar,19.Contreras M.P. al.Sensor helpers pathogens.EMBO 42e111519PubMed messengers, calcium ion derivations. crosstalk between negative effectors host regulators. directly indirectly. For example, interaction benthamiana TNL Roq1 (recognition XopQ its recognized (Xanthomonas outer Q) [20.Schultink al.Roq1 Xanthomonas Pseudomonas HopQ1.Plant 92: 787-795Crossref (82) Scholar,21.Martin R. al.Structure ROQ1 recognizing XopQ.Science. 370eabd9993Crossref (174) Scholar], arabidopsis RPP1 Peronospora parasitica ATR1 (Arabidopsis thaliana [22.Ma al.Direct pathogen-induced complex holoenzyme.Science. 370abe3069Crossref (169) Scholar,23.Krasileva K.V. al.Activation Arabidopsis specified planta association cognate oomycete effector.Plant 2010; 22: 2444-2458Crossref (220) wheat Sr35 (stem rust 35) AvrSr35 [24.Zhao Y.-B. al.Pathogen triggers mechanism.Sci. Adv. 8eabq5108Crossref (13) 25.Forderer al.A defines common principles channels.Nature. 610: 532-539Crossref 26.Salcedo al.Variation determines stem race Ug99.Science. 358: 1604-1606Crossref (117) spp, Hyaloperonospora parasitica, Puccinia graminis tritici, respectively. monitoring effector-mediated perturbations targets. example ZAR1 (HOPZ-ACTIVATED RESISTANCE [27.Lewis al.Allele-specific attenuation syringae HopZ1a type III protein.PLoS Genet. 6e1000894Crossref (111) exists preformed kinase RKS1 (resistance-related normal conditions recognizes AvrAC indirectly uridylylation another PBL2 (PBS1-like 2) [28.Wang al.Ligand-triggered allosteric ADP release primes complex.Science. 364aav5868Crossref (231) Scholar,29.Wang G. al.The decoy substrate pseudokinase specify modified-self plants.Cell Host Microbe. 2015; 18: 285-295Abstract (156) Scholar]; AvrAC-uridylylated associates consequently activates ZAR1-mediated immunity. Alternatively, cases, may not necessarily CHS3 (CHILLING SENSITIVE 3)/CSA1 (CONSTITUTIVE SHADE-AVOIDANCE pair detects PRR coreceptor BAK1 (BRASSINOSTEROID INSENSITIVE 1-associated [30.Yang al.Allelic variation CHS3/CSA1 reveals functional cell-death regulatory modes.Cell 30: 1701-1716Abstract (7) malectin-like receptor-like LET1 autoimmunity SUMM2 (SUPPRESSOR OF MKK1 MKK2) MEKK2 (MAP/ERK kinase-2) scaffolding [31.Liu LETUM1 modulates scaffolding.Nat. Plants. 6: 1106-1115Crossref (26) Effector leads termed 1). Cryoelectron microscopy (cryoEM) analyses show containing ZAR1, RKS1, uridylylated (PBL2UMP) forms wheel-like [32.Wang al.Reconstitution structure conferring immunity.Science. 364aav5870Crossref (368) 1, left). Pentamerization mainly NOD module PBL2UMP being presented rim wheel 2). contrast AvrSr35, remarkably similar Scholar,25.Forderer more study showed oligomeric NRG1 likely formed plasma membrane (PM) [33.Feehan al.Oligomerization requires activation.Proc. 120e2210406120Crossref (3) Interestingly, resistosomes, underlying await further elucidation By comparison, end results right). induced [21.Martin As seen primarily Structural modeling sequestered inactive Scholar,24.Zhao Scholar,32.Wang suggesting oligomerization. it remains unknown whether all resistosomes. Many pairs, executor initiating [34.Xi al.Insight mode action paired receptors.Essays 513-526Crossref (2) Whether how represent challenge understanding mechanisms. cryoEM resistosome, five α1-helices solvent-exposed shaped like channel pore Functional data support ETI. Importantly, electrophysiological evidence supports [35.Bi calcium-permeable triggering 184: 3528-3541Abstract (181) bottom Single molecule imaging hours before loss PM integrity These suggest acts α1 distantly related [36.Adachi al.An motif functionally across species.eLife. 8e49956Crossref (92) Ca2+-channel among Indeed, bears [25.Forderer essential helix well-defined resistosome. It environment funnel-shaped localization shown RPM1 (resistance pv. maculicola [37.El Kasmi F. al.Signaling plasma-membrane self-association full-length protein.Proc. E7385-E7394Crossref (70) RPS2 P. [38.Axtell M.J. Staskawicz B.J. Initiation RPS2-specified coupled AvrRpt2-directed elimination RIN4.Cell. 2003; 112: 369-377Abstract (643) bethamiana Tm-22 [39.Wang NB-ARC domain-mediated CC domain.PLoS Pathog. 16e1008475Crossref (20) ZAR1-like examined. TNL-activated hNLRs, NRG1s, ADR1s display autoactive Scholar,40.Jacob "helper" nonselective cation channels.Science. 373: 420-425Crossref (125) indicating Solanaceae (SD) found noncanonical [41.Seong K. al.Evolution genes wild tomato species.New 227: 1530-1543Crossref (36) themselves structures. undetermined activity. Multiple lines extracellular [42.Xu tale families: channels immunity.Plant 1551-1567Crossref (19) Scholar,43.Kim N.H. al.Con-Ca2+-tenating channels.New 234: 813-818Crossref (8) Elevation concentrations one earliest during Gain-of-function mutations CNGC19/20 (CYCLIC NUCLEOTIDE GATED CHANNEL19/20) increased constitutively (enhanced susceptibility 1)- SA (salicylic acid)-dependent [44.Yu X. kinases BAK1/SERK4 channel-mediated cellular homeostasis containment.Curr. 29: 3778-3790Abstract (64) Scholar,45.Zhao mis-regulated cyclic nucleotide-gated cytosolic elevation Arabidopsis.New 230: 1078-1094Crossref (33) Unregulated Sr35, NRG1, ADR1 sufficient recapitulate CNL-mediated eukaryotic Although released internal pools contribute pharmacological blocking compartments ruthenium red (RR) less efficient inhibition HR than blocker LaCl3 [46.Gao heat-activated 6 involved heat shock responses.Plant 2012; 70: 1056-1069Crossref supporting notion major responses. Nuclear [47.Ludke D. yet? Nucleocytoplasmic coordination immunity.New 236: 24-42Crossref mechanism associated enigmatic. Several interact factors [48.Wang al.Diversity, receptors.J. Integr. 63: 283-296Crossref (11) programming nucleus. transcription set genes, other reprogramming. NRG1A PM- nucleus-localized activation, only PM-resident oligomers oligomerization-independent function. Increases nuclear free reported various stresses [49.Pauly nucleus together cytosol generates specific signatures tobacco suspension culture cells.Cell Calcium. 2001; 413-421Crossref 50.van Der Luit A.H. al.Distinct calmodulin expression tobacco.Plant Physiol. 1999; 121: 705-714Crossref 51.Xiong T.C. al.Isolated nuclei mechanical thermal sensors signalling.Plant 2004; 40: 12-21Crossref should kept mind spatial distribution uniform, steep gradients few nanometers away [52.Pangršič al.EF-hand buffers exocytosis sensory hair cells.Proc. E1028-E1037Crossref transported PM-localized and/or reach possible continuum organelle endoplasmic reticulum (ER). Such result An CNGC15 symbiotic oscillation [53.Charpentier al.Nuclear-localized oscillations.Science. 352: 1102-1105Crossref While signals, they TNLs, CNLs, NADases. ATR1-induced significantly enhances activity, holoenzyme enzymatic encoded TIR [54.Wan L. al.TIR NAD+-cleaving enzymes promote death.Science. 365: 799-803Crossref (226) Scholar,55.Horsefield al.NAD+ cleavage animal pathways.Science. 793-799Crossref (244) TIR-only bacterial HopBA1 (RBA1) [56.Nishimura M.T. al.TIR-only RBA1 Arabidopsis.Proc. E2053-E2062Crossref (93) Tetramerization composite sites Scholar,22.Ma [57.Shi SARM1 recognition, small molecules.Mol. 82: 1643-1659Abstract (27) 58.Manik M.K. al.Cyclic ribose isomers: production, chemical structures, signaling.Science. 377eadc8969Crossref 59.Hogrel nucleotide-induced helical effector.Nature. 608: 808-812Crossref analogs PAD4 (phytoalexin deficient 4)

Language: Английский

---

Conservation and similarity of bacterial and eukaryotic innate immunity

Nature Reviews Microbiology, Journal Year: 2024, Volume and Issue: 22(7), P. 420 - 434

Published: Feb. 28, 2024

Language: Английский

---

Oligomerization-mediated autoinhibition and cofactor binding of a plant NLR

Nature, Journal Year: 2024, Volume and Issue: 632(8026), P. 869 - 876

Published: June 12, 2024

Abstract Nucleotide-binding leucine-rich repeat (NLR) proteins play a pivotal role in plant immunity by recognizing pathogen effectors 1,2 . Maintaining balanced immune response is crucial, as excessive NLR expression can lead to unintended autoimmunity 3,4 Unlike most NLRs, the required for cell death 2 (NRC2) belongs small group characterized constitutively high without self-activation 5 The mechanisms underlying NRC2 autoinhibition and activation are not yet understood. Here we show that Solanum lycopersicum (tomato) ( Sl NRC2) forms dimers tetramers higher-order oligomers at elevated concentrations. Cryo-electron microscopy shows an inactive conformation of these oligomers. Dimerization oligomerization only stabilize state but also sequester from assembling into active form. Mutations dimeric or interdimeric interfaces enhance pathogen-induced Nicotiana benthamiana cryo-electron structures unexpectedly inositol hexakisphosphate (IP 6 ) pentakisphosphate bound inner surface C-terminal domain NRC2, confirmed mass spectrometry. phosphate-binding site impair phosphate binding NRC2-mediated N. Our study indicates negative regulatory mechanism suggests phosphates cofactors NRCs.

Language: Английский

---

Plant NLR immunity activation and execution: a biochemical perspective

Open Biology, Journal Year: 2024, Volume and Issue: 14(1)

Published: Jan. 1, 2024

Plants deploy cell-surface and intracellular receptors to detect pathogen attack trigger innate immune responses. Inside host cells, families of nucleotide-binding/leucine-rich repeat (NLR) proteins serve as sensors or downstream mediators defence outputs cell death, which prevent disease. Established genetic underpinnings NLR-mediated immunity revealed various strategies plants adopt combat rapidly evolving microbial pathogens. The molecular mechanisms NLR activation signal transmission components controlling execution were less clear. Here, we review recent protein structural biochemical insights plant sensor signalling functions. When put together, the data show how different families, whether transducers, converge on nucleotide-based second messengers cellular calcium confer immunity. Although pathogen-activated NLRs in engage plant-specific machineries promote defence, comparisons with mammalian receptor counterparts highlight some shared working principles for across kingdoms.

Language: Английский

---

Substrate-induced condensation activates plant TIR domain proteins

Nature, Journal Year: 2024, Volume and Issue: 627(8005), P. 847 - 853

Published: March 13, 2024

Abstract Plant nucleotide-binding leucine-rich repeat (NLR) immune receptors with an N-terminal Toll/interleukin-1 receptor (TIR) domain mediate recognition of strain-specific pathogen effectors, typically via their C-terminal ligand-sensing domains 1 . Effector binding enables TIR-encoded enzymatic activities that are required for TIR–NLR (TNL)-mediated immunity 2,3 Many truncated TNL proteins lack effector-sensing but retain similar and 4,5 The mechanism underlying the activation these TIR remain unclear. Here we show substrates NAD + ATP induces phase separation in vitro. A condensation occurs a protein expressed its native promoter response to inoculation planta. formation condensates is mediated by conserved self-association interfaces predicted intrinsically disordered loop region TIRs. Mutations disrupt impair cell death activity proteins. Our data reveal as provide insight into substrate-induced autonomous signalling confer plant immunity.

Language: Английский

---

Activation of the helper NRC4 immune receptor forms a hexameric resistosome

Cell, Journal Year: 2024, Volume and Issue: 187(18), P. 4877 - 4889.e15

Published: Aug. 1, 2024

Language: Английский

---

TIR signaling activates caspase-like immunity in bacteria

Science, Journal Year: 2025, Volume and Issue: 387(6733), P. 510 - 516

Published: Jan. 30, 2025

Caspase family proteases and Toll/interleukin-1 receptor (TIR)-domain proteins have central roles in innate immunity regulated cell death humans. We describe a bacterial immune system comprising both caspase-like protease TIR-domain protein. found that the TIR protein, once it recognizes phage invasion, produces previously unknown signaling molecule adenosine 5′-diphosphate-cyclo[N7:1′′]-ribose (N7-cADPR). This specifically activates protease, which then indiscriminately degrades cellular to halt replication. The TIR-caspase defense system, we denote as type IV Thoeris, is abundant bacteria efficiently protects against propagation. Our study highlights diversity of TIR-produced molecules demonstrates by caspase an ancient mechanism immunity.

Language: Английский

---

A rare PRIMER cell state in plant immunity

Nature, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 8, 2025

Abstract Plants lack specialized and mobile immune cells. Consequently, any cell type that encounters pathogens must mount responses communicate with surrounding cells for successful defence. However, the diversity, spatial organization function of cellular states in pathogen-infected plants are poorly understood 1 . Here we infect Arabidopsis thaliana leaves bacterial trigger or supress integrate time-resolved single-cell transcriptomic, epigenomic transcriptomic data to identify states. We describe cell-state-specific gene-regulatory logic involves transcription factors, putative cis -regulatory elements target genes associated disease immunity. show a rare population emerges at nexus immune-active hotspots, which designate as primary responder (PRIMER) PRIMER have non-canonical signatures, exemplified by expression genome accessibility previously uncharacterized factor, GT-3A, contributes plant immunity against pathogens. surrounded another state (bystander) activates long-distance cell-to-cell signalling. Together, our findings suggest interactions between these propagate across leaf. Our molecularly defined spatiotemporal atlas provides functional regulatory insights into plants.

Language: Английский

---

Plant pathogenic fungi hijack phosphate signaling with conserved enzymatic effectors

Science, Journal Year: 2025, Volume and Issue: 387(6737), P. 955 - 962

Published: Feb. 27, 2025

Inorganic phosphate (Pi) is essential for life, and plant cells monitor Pi availability by sensing inositol pyrophosphate (PP-InsP) levels. In this work, we describe the hijacking of a conserved family Nudix hydrolase effectors from pathogenic Magnaporthe Colletotrichum fungi. Structural enzymatic analyses effector demonstrate that they selectively hydrolyze PP-InsP. Gene deletion experiments in oryzae, higginsianum, graminicola indicate PP-InsP hydrolysis substantially enhances disease symptoms diverse pathosystems. Further, show induces starvation signaling plants. Our study elucidates molecular mechanism, used multiple phytopathogenic fungi, manipulates highly pathway to exacerbate disease.

Language: Английский

---