Biology subjects

Spoel, S. H.

Publications and source records attributed to Spoel, S. H..

11 recordsLinked to original sources

bioRxiv2026

HECT-type ligases facilitate autoubiquitination and degradation of other ubiquitin ligases to activate plant immunity

The ubiquitin-proteasome system (UPS) serves as the primary proteolytic machinery in eukaryotes, governing intracellular protein turnover to maintain proteome homeostasis. In plants, the HECT-type UPL3/4 ubiquitin ligases play vital roles in developmental and immune signaling. After ubiquitination by pathway-specific E3 ligases, substrates are physically relayed to proteasome-associated UPL3/4 ligases for further modification, which is necessary for their proteasome-mediated degradation. In this study, we investigated if the cellular influence of UPL3/4 extends beyond their direct role in substrate degradation. We discovered that UPL3/4 govern the ubiquitination not only of a broad array of immune-related substrates, but also of many UPS components, including E3 ligases. UPL3 physically interacts with PUB22, a pathway-specific U-box E3 ligase that negatively regulates immunity. PUB22 is controlled by a phospho-switch that converts it from an instable autoubiquitinated state to a stable phosphorylated E3 ligase that marks substrates for degradation. Remarkably, UPL3 only interacted with unphosphorylated PUB22 and facilitated its autoubiquitination-mediated degradation, thereby promoting the accumulation of PUB22 substrates. Moreover, the compromised immune phenotypes of upl3 upl4 mutant plants were largely dependent on PUB22 and its close paralogues. Thus, UPL3/4 control the stability of immune-related substrates not only through direct ubiquitination, but also indirectly by promoting autoubiquitination of PUB22 ligase and its paralogues. Controlling the stability of autoubiquitinating E3 ligases may be a universal mechanism whereby HECT-type ligases and the proteasomes they associated with, orchestrate cellular proteostasis in eukaryotes. Significance StatementThe ubiquitin-proteasome system (UPS) governs intracellular protein turnover to maintain proteome homeostasis in eukaryotes. Proteasome-associate HECT-type ubiquitin ligases play an important role in processing and degrading substrates delivered to the proteasome by pathway-specific E3 ligases. Here, we discover that in plants, HECT-type ligases not only promote the degradation of substrates, they also modify the E3 ligases that target these substrates to the proteasome. Specifically, HECT-type ligases facilitated or expanded the autoubiquitination of immune-suppressive E3 ligases, resulting in their proteasome-mediated degradation and onset of immunity. Our discoveries suggest that during plant immunity, HECT-type ligases and the proteasomes they associate with, control cellular proteostasis by governing the stabilities of both E3 ligases and their substrates.

plant biology↗

bioRxiv2026

GSNOR-dependent nitric oxide homeostasis promotes recovery from repeated climate stress across generations in Arabidopsis thaliana

Climate change exposes plants to recurrent and interacting stresses, yet the extent to which these effects persist across generations, and the mechanisms involved, remain unclear. We propagated Arabidopsis thaliana wild type (WT, Col-0) and nitric oxide homeostasis mutant gsnor1-3 (hereafter, gsnor-ko) for five successive generations. Plants were grown under control, drought, elevated CO2, O3, warm temperature, and combined treatment scenarios for the first three generations (G1-G3), followed by two recovery generations under control conditions (G4-G5). We quantified rosette growth, photosynthetic traits, seed production, and transcriptome dynamics by RNA-seq. Across environments, gsnor-ko showed reduced vegetative growth and reproductive output relative to WT. Transcriptomic responses were strongly scenario- and generation-dependent, with the largest differential expression shifts observed under warm-climate and combined-treatment conditions. Compared with WT, gsnor-ko displayed broader gene overlap across generations and stronger retention or reconfiguration of stress-responsive states after stress withdrawal. Functional enrichment and candidate-gene analyses identified pathways/components linked to DNA methylation, heterochromatin maintenance, histone ubiquitination, m6A RNA regulation, and methyl-donor metabolism. Together, these results support a model in which GSNOR activity promotes transcriptomic recovery after repeated climate stress, whereas impaired GSNOR function shifts responses toward multi-generational persistence and epigenetically associated regulatory reconfiguration. HighlightGSNOR-dependent nitric oxide homeostasis promotes transcriptomic resetting after repeated climate stress, whereas impaired NO homeostasis favours multigenerational persistence and chromatin- and RNA-linked regulatory reconfiguration. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/742973v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@6031c6org.highwire.dtl.DTLVardef@163be5borg.highwire.dtl.DTLVardef@1665e9eorg.highwire.dtl.DTLVardef@1cde833_HPS_FORMAT_FIGEXP M_FIG C_FIG

plant biology↗

bioRxiv2026

Ubiquitin recognition integrates plant immune signaling by cell-surface and intracellular receptors

Plant immunity is activated by cell-surface and intracellular receptors that detect pathogen-derived molecules. Mutual potentiation between these two receptor types is essential for robust disease resistance, but the mechanisms underpinning integrated dual receptor immunity remain unknown. Here, we found that activation of the intracellular receptor, ZAR1, induces its site-specific modification by non-proteolytic ubiquitin chains. ZAR1-anchored ubiquitin chains promote oligomerization of ZAR1 into a calcium-permeable resistosome pore and are recognized by RH3, a ubiquitin-binding DEAD-box RNA helicase. Remarkably, RH3 recruits both ZAR1 resistosomes and cell-surface receptor components into a dual receptor complex, thereby enhancing calcium-dependent mRNA translation of core defense proteins and establishing robust immunity. These findings identify ubiquitin recognition as the missing link in mutual potentiation of plant immunity by cell-surface and intracellular receptors.

plant biology↗

bioRxiv2025

TGA family of transcription factors are nitric oxide sensors in the root stem cell niche

Plant root development is tightly regulated and depends on the coordination among different cell types located in the root stem cell niche (SCN). A complex network of regulators includes the gasotransmitter nitric oxide (NO) known to inhibit primary root growth, modify root architecture and alter meristem organisation. However, little is known about the molecular targets and the mechanistic insights of this signalling molecule during root SCN homeostasis. Here, genetic, molecular and physiological evidence support that TGA transcription factors of the bZIP family, and specifically PERIANTHIA (PAN), contribute to NO-mediated root stem cell regulation. NO induces reversible S-nitrosylation of PAN, thereby inhibiting its binding to DNA and associated transcriptional activation. In turn, the protein-SNO reductase activity of TRXh5 is able to denitrosylate PAN. This switch on and off mechanism of PAN activation modulates WOX5 expression and quiescent centre activity, which alters columella stem cell maintenance through a NO-dependent mechanism. Different TGA members interact with PAN and are also S-nitrosylated, whose expression in the root SCN together with their accumulated mutations contribute to NO sensing during root apical meristem development. Collectively, these findings reveal a molecular framework where NO controls the ability of PAN to bind to its targets as a master regulator of the root SCN.

plant biology↗

bioRxiv2025

TOC1 supresses PAMP-triggered immunity in Arabidopsis

The circadian clock synchronizes plants with the rhythmic changes in their environment created by the cycle of day and night. Plant-pathogen interactions are influenced by this rhythmic cycle and a functioning circadian clock is essential for eQective resistance to pathogens. In this study we investigate the relationship between PAMP-triggered immune signalling and the circadian clock in Arabidopsis. We found that early PAMP-triggered immune responses including flg22- induced FRK1 transcript levels and the flg22-induced ROS burst are enhanced at subjective dusk rather than subjective dawn. Overexpression of clock gene TOC1 supressed these defence responses to flg22 and resulted in increased susceptibility to Pseudomonas syringae. Additionally, treatment with flg22 and elf18 altered the rhythmicity of CCA1 and TOC1. These results contribute to the currently small amount of research investigating the interactions between the circadian clock and PAMP-triggered immunity. Importantly, we establish the detrimental eQect of TOC1 overexpression on PAMP-triggered defence responses. Further investigation into how clock components regulate early immune responses is essential for improving our understanding of plant health.

plant biology↗

bioRxiv2025

Mobile immune signals potentiate salicylic acid-mediated plant immunity via WRKY38/62 transcription factors

Systemic acquired resistance (SAR) is a broad-spectrum plant immune response that provides protection against various pathogens. Activation of SAR requires mobile immune signals as well as the indispensable immune hormone salicylic acid (SA). Nonetheless, it remains unknown how mobile signals integrate with the SA signal to produce functional SAR responses. Here, we demonstrate that the mobile signals, azelaic acid (AzA) and N-hydroxy-pipecolic acid (NHP), respectively dampened and potentiated SA-induced transcriptional reprogramming of thousands of genes. Indeed, NHP enhanced stability of the SA receptor protein NPR1, and unlike AzA, it dramatically increased the effectiveness of SA-induced immunity against bacterial infection by 10-fold. Analysis of NHP-primed, SA-responsive gene promoters indicated that WRKY transcription factors play an important role in integrating these two immune signals. While responsiveness to SA remained largely unaffected by mutation of WRKY38 and WRKY62, it abolished NHP-mediated potentiation of SA-induced gene expression and immunity. Collectively, our findings reveal mobile signals potentiate SA-mediated plant immunity via WRKY38/62 transcription factors. TeaserActivation of broad-spectrum immunity in plants requires mutual potentiation between multiple distinct immune signals.

plant biology↗

bioRxiv2023

The pathogenic coronatine toxin hijacks host redox signalling to suppress plant immunity

Reciprocal antagonism between the hormones salicylic acid (SA) and jasmonic acid (JA) is particularly important during infection by the bacterial pathogen Pseudomonas syringae. P. syringae secretion of the virulence-promoting toxin, coronatine, is thought to suppress SA-induced immune responses by manipulating host JA signalling. Here, we report an unexpected JA-independent role of coronatine in promoting pathogen virulence. While JA induced resistance to P. syringae, coronatine promoted pathogen virulence by suppressing cellular accumulation of glutathione, a vital antioxidant required for immunity. Coronatine-mediated suppression of glutathione levels prevented activation of NPR1, a redox-sensitive master regulator of SA-responsive immune genes. Moreover, the accrual of nitric oxide (NO) restored virulence of coronatine-deficient P. syringae, but was counteracted by expression of the host S-nitrosothiol reductase, Thioredoxin h5. Thus, our findings indicate P. syringae utilises coronatine to suppresses host immunity by precisely manipulating glutathione- and NO-mediated redox signalling networks.

plant biology↗

bioRxiv2023

Crosstalk between salicylic acid signaling and the circadian clock promotes an effective immune response

The rotation of Earth creates a cycle of day and night, leading to predictable changes in environmental conditions. The circadian clock synchronizes an organism with these environmental changes and alters their physiology in anticipation. Prediction of the probable timing of pathogen infection enables plants to prime their immune system without wasting resources or sacrificing growth. Here, we explore the relationship between the immune hormone salicylic acid (SA), and the circadian clock in Arabidopsis. We found that SA altered circadian rhythmicity through the SA receptor and master transcriptional coactivator, NPR1. Reciprocally, the circadian clock gates SA-induced expression of NPR1-dependent immune genes. Furthermore, the clock gene CCA1 is essential for SA-induced immunity to the major bacterial plant pathogen Pseudomonas syringae. These results reveal new interactions between the circadian clock and SA signaling which produce an effective immune response. Understanding how and why the immune response in plants is linked to the circadian clock is crucial in working towards improved crop productivity.

plant biology↗

bioRxiv2021

Proteasome-associated ubiquitin ligase relays target plant hormone-specific transcriptional activators

The ubiquitin-proteasome system is vital to hormone-mediated developmental and stress responses in plants. Ubiquitin ligases target hormone-specific transcriptional activators (TAs) for degradation, but how TAs are processed by proteasomes remains unknown. We report that in Arabidopsis the salicylic acid-and ethylene-responsive TAs, NPR1 and EIN3, are relayed from pathway-specific ubiquitin ligases to proteasome-associated HECT-type UPL3/4 ligases. Activity and stability of NPR1 was regulated by sequential action of three ubiquitin ligases, including UPL3/4, while proteasome processing of EIN3 required physical handover between ethylene-responsive SCFEBF2 and UPL3/4 ligases. Consequently, UPL3/4 controlled extensive hormone-induced developmental and stress-responsive transcriptional programmes. Thus, our findings identify unknown ubiquitin ligase relays that terminate with proteasome-associated HECT-type ligases, which may be a universal mechanism for processive degradation of proteasome-targeted TAs and other substrates. One-Sentence SummaryTranscriptional activators are targeted by proteasomal ubiquitin ligase relays that control their activity and stability.

plant biology↗

bioRxiv2021

Mechanosensory trichome cells evoke a mechanical stimuli-induced immune response in plants

Perception of pathogen-derived ligands by corresponding host receptors is a pivotal strategy in eukaryotic innate immunity. In plants, this is complemented by circadian anticipation of infection timing, promoting basal resistance even in the absence of pathogen threat. Here, we report that trichomes, hair-like structures on the epidermis, directly sense external mechanical forces caused by raindrops to anticipate waterborne infections in Arabidopsis thaliana. Exposure of leaf surfaces to mechanical stimuli initiates the concentric propagation of intercellular calcium waves away from trichomes to induce defence-related genes. Propagating calcium waves enable effective immunity against pathogenic microbes through the calmodulin-binding transcription activator 3 (CAMTA3) and mitogen-activated protein kinases. We propose a novel layer of plant immunity in which trichomes function as mechanosensory cells to detect potential risks.

plant biology↗

bioRxiv2019

Dynamic ubiquitination determines transcriptional activity of the plant immune coactivator NPR1

Activation of systemic acquired resistance in plants is associated with transcriptome reprogramming induced by the unstable coactivator NPR1. Immune-induced ubiquitination and proteasomal degradation of NPR1 are thought to facilitate continuous delivery of active NPR1 to target promoters, thereby maximising gene expression. Because of this potentially costly sacrificial process, we investigated if ubiquitination of NPR1 plays transcriptional roles prior to its proteasomal turnover. Here we show ubiquitination of NPR1 is a processive event in which initial modification by a Cullin-RING E3 ligase promotes its chromatin association and expression of target genes. Only when polyubiquitination of NPR1 is enhanced by the E4 ligase, UBE4, it is targeted for proteasomal degradation. Conversely, ubiquitin ligase activities are opposed by UBP6/7, two proteasome-associated deubiquitinases that enhance NPR1 longevity. Thus, immune-induced transcriptome reprogramming requires sequential actions of E3 and E4 ligases balanced by opposing deubiquitinases that fine-tune activity of NPR1 without strict requirement for its sacrificial turnover.

plant biology↗