Single-molecule analysis reveals the phosphorylation of FLS2 regulates its spatiotemporal dynamics and immunity

作者：Yaning Cui, Hongping Qian, Jinhuan Yin, Changwen Xu, Pengyun Luo, Xi Zhang, Meng Yu, Bodan Su, Xiaojuan Li, Jinxing Lin · 发表于：eLife · 年份：2024 · DOI：10.7554/elife.91072.2 · 被引用次数：1 · 研究领域：Glycosylation and Glycoproteins Research、Carbohydrate Chemistry and Synthesis

Summary Phosphorylation of receptor kinase (RK) is pivotal for signaling in pattern-triggered immunity (PTI). The Arabidopsis thaliana FLAGELLIN-SENSITIVE2 (FLS2) is a conserved 22 amino acid sequence in the N-terminal region of flagellin (flg22), initiating plant defense pathways. However, the dynamic FLS2 phosphorylation regulation at the plasma membrane in response to flg22 needs further elucidation. Through single-particle tracking, we demonstrated that the Ser-938 phosphorylation site influences flg22-induced FLS2 spatiotemporal dynamics and dwell time. Förster resonance energy transfer-fluorescence lifetime (FRET-FLIM) imaging microscopy, coupled with protein proximity indexes (PPI), revealed increased co-localization of FLS2/FLS2S938D-GFP with AtRem1.3-mCherry in response to flg22. In contrast, FLS2S938A-GFP shows no significant changes, indicating that Ser-938 phosphorylation influences the efficient FLS2 sorting into AtRem1.3-associated microdomains. Significantly, Ser-938 phosphorylation enhanced flg22-induced internalization and immune responses, thus demonstrating its regulatory role in FLS2 partitioning into functional AtRem1.3-associated microdomains for activating flg22-induced plant immunity.