Pupylation-based proximity labeling reveals APC/C-mediated ubiquitination and degradation of PEROXIN11C in soybean abiotic stress responses by Yue Zhang, Xinxin Yang, Tong Chen, Kaidi Su, Yuhong Zhu, Shuo Li, Fengning Xiang, and Qing Lu ISBN 10.1016/J.CELREP.2025.116331 instant download

Pupylation-based proximity labeling reveals APC/C-mediated ubiquitination and degradation of PEROXIN11C in soybean abiotic stress responses by Yue Zhang, Xinxin Yang, Tong Chen, Kaidi Su, Yuhong Zhu, Shuo Li, Fengning Xiang, and Qing Lu ISBN 10.1016/J.CELREP.2025.116331 instant download

SUMMARY: Proximity-dependent labeling coupled with mass spectrometry (PL-MS) is a powerful approach for mapping low-affinity, transient protein-protein interactions (PPIs). While widely employed in animal systems, its application in plants has been restricted primarily to TurboID/miniTurbo-based systems due to the limited enzyme compatibility. To address this, we optimized the pupylation-based interaction tagging (Pup-IT) with a dexamethasone-inducible expression strategy in Nicotiana benthamiana and soybean (Glycine max). Using PupIT, we mapped the interactome of the soybean anaphase-promoting complex/cyclosome (APC/C) and identified the peroxisomal membrane protein GmPEX11C as a substrate. GmAPC/C ubiquitinates GmPEX11C atlysine 116, targeting it for proteasomal degradation. In addition, Gmilpa1/apc8 mutant exhibited enhanced salt and drought tolerance, whereas knockdown of GmPEX11C via RNA interference abolishes this stress-resistant phenotype. Our study establishes Pup-IT as a useful tool for interrogating PPIs in planta and uncovers a mechanistic link between APC/C-mediated ubiquitination and abiotic stress adaptation in soybean,offering potential avenues for crop improvement.