Split-YFP-coupled interaction-dependent TurboID identifies new functions of basal cell polarity in Arabidopsis

• 成果类型: Article
• 署名作者: Huang, Aobo; Zhang, Jinming; Liu, Zhendong; Schoen, Vanessa; Verma, Deepanjali; Zheng, Haiyan; Pedmale, Ullas, V; Dong, Juan
• 署名单位: Rutgers University System; Rutgers University New Brunswick; Texas A&M University System; Texas A&M University College Station; Cold Spring Harbor Laboratory; Rutgers University System; Rutgers University New Brunswick; Rutgers University Biomedical & Health Sciences; Rutgers University System; Rutgers University New Brunswick; Rutgers University System; Rutgers University New Brunswick
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-15231
• DOI: 10.1073/pnas.2502445122
• 发表日期: 2025-08-12
• 关键词: membrane-associated protein; planar polarity; auxin transport; stomatal lineage; agcviii kinases; exocyst complex; division; GROWTH; light; SYSTEM

摘要

The formation of a body axis is one of the fundamental steps in developmental patterning in multicellular organisms. Ectopic expression of the stomatal protein BASL (BREAKING OF ASYMMETRY IN THE STOMATAL LINEAGE) reveals a proximal-distal cell polarity field in the leaf and an apical-basal field in the hypocotyl and root of Arabidopsis. This provides a framework for uncovering molecular components of body-axis cell polarity in higher plants. In this study, we developed a proximity labeling-based complementation system, termed tSYID (tandem fusion of Split-YFP and TurboID), to enable simultaneous visualization and identification of the tissue-wide cell polarity module marked by interacting BASL and BREVIS RADIX-LIKE 2 (BRXL2) proteins in Arabidopsis. This contact-based tSYID system focuses on identifying the protein interactome proximal to the protein-protein interaction site. By combining proximity labeling and mass spectrometry, we identified protein candidates associated with the tSYID-BASL/BRXL2 module, many of which remain uncharacterized in plants. Among them, an AGC protein kinase displayed highly polarized localization at the basal membrane in the hypocotyl and root. Knocking out two close AGChomologs in Arabidopsis resulted in compromised hypocotyl growth in the dark, suggesting a functional connection between basal cell polarity and plant upward growth. Our study demonstrates the power of using the Split-TurboID strategy to uncover proximal proteomes near protein activity sites and highlights an underappreciated crosstalk between developmentally programmed body axes and environmentally influenced growth vectors.