ABSTRACT
Plant pattern-recognition receptors perceive microorganism-associated molecular patterns to activate immune signalling1,2. Activation of the pattern-recognition receptor kinase CERK1 is essential for immunity, but tight inhibition of receptor kinases in the absence of pathogen is crucial to prevent autoimmunity3,4. Here we find that the U-box ubiquitin E3 ligase OsCIE1 acts as a molecular brake to inhibit OsCERK1 in rice. During homeostasis, OsCIE1 ubiquitinates OsCERK1, reducing its kinase activity. In the presence of the microorganism-associated molecular pattern chitin, active OsCERK1 phosphorylates OsCIE1 and blocks its E3 ligase activity, thus releasing the brake and promoting immunity. Phosphorylation of a serine within the U-box of OsCIE1 prevents its interaction with E2 ubiquitin-conjugating enzymes and serves as a phosphorylation switch. This phosphorylation site is conserved in E3 ligases from plants to animals. Our work identifies a ligand-released brake that enables dynamic immune regulation.
Subject(s)
Oryza , Plant Immunity , Plant Proteins , Ubiquitin , Animals , Chitin/metabolism , Homeostasis , Ligands , Oryza/enzymology , Oryza/immunology , Oryza/metabolism , Oryza/microbiology , Phosphorylation , Plant Proteins/antagonists & inhibitors , Plant Proteins/immunology , Plant Proteins/metabolism , Ubiquitin/metabolism , Ubiquitin-Conjugating Enzymes/metabolism , Ubiquitin-Protein Ligases/antagonists & inhibitors , Ubiquitin-Protein Ligases/chemistry , Ubiquitin-Protein Ligases/metabolism , Ubiquitination , Phosphoserine/metabolism , Conserved SequenceABSTRACT
In this study, crude polysaccharide (LAG-C) and homogeneous arabinogalactan (LAG-W) were isolated from Qinling Larix kaempferi of Shaanxi Province. Bioactivity assays showed that LAG-W and LAG-C enhanced the phagocytic ability, NO secretion, acid phosphatase activity, and cytokine production (IL-6, IL-1ß, and TNF-α) of RAW264.7 macrophages. Notably, LAG-W exhibited a significantly stronger immunomodulatory effect than LAG-C. The primary structure of LAG-W was characterised by chemical methods (monosaccharide composition, methylation analysis, and alkali treatment) and spectroscopic techniques (gas chromatography-mass spectrometry, high-performance liquid chromatography-mass spectrometry, and 1D/2D nuclear magnetic resonance). LAG-W was identified as a 22.08 kilodaltons (kDa) neutral polysaccharide composed of arabinose and galactose at a 1:7.5 molar ratio. Its backbone consisted of repeated â3)-ß-Galp-(1â residues. Side chains, connected at the O-6 position, were mainly composed of T-ß-Galp-(1â and T-ß-Galp-(1â6)-ß-Galp-(1â residues. And it also contained small amounts of T-ß-Arap-(1â, T-α-Araf-(1â6)-ß-Galp-(1â6)-ß-Galp-(1â, and T-α-Araf-(1â3)-α-Araf-(1â6)-ß-Galp-(1â residues. By structurally and functionally characterising L. kaempferi polysaccharides, this study opens the way for the valorisation of this species.