CLE19 suppresses brassinosteroid signaling output via the BSL-BIN2 module to maintain BES1 activity and pollen exine patterning in Arabidopsis

doi:10.1111/jipb.70024

• Research Article •

CLE19 suppresses brassinosteroid signaling output via the BSL-BIN2 module to maintain BES1 activity and pollen exine patterning in Arabidopsis

Shuangshuang Wang^1†,‡, Shiting Zhang^1†, Ying Yu^1†, Jianzheng Wang¹, Jingya Wang¹, Mengyu Li¹, Jianan Lu¹, Juanying Ye¹, Hanji Li¹, Yeqiao Liu¹, Yuhan Zhao¹, Wen Song³, Juan Dong^4,5, Jia Li⁶, Chunming Liu⁷, Hong Ma², Fang Chang^1*

1. State Key Laboratory of Genetics and Development of Complex Phenotypes, School of Life Sciences, Fudan University, Shanghai 200438, China

2. Department of Biology, the Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA

3. State Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing 100193, China

4. The Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA

5. Department of Plant Biology, Rutgers, the State University of New Jersey, New Brunswick, NJ 08901, USA

6. Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design, School of Life Sciences, Guangzhou University, Guangzhou 510006, China

7. School of Advanced Agricultural Sciences, Peking University, Beijing 100871, China

^†These authors contributed equally to this work.

^‡Present address: School of Life Sciences, East China Normal University, Shanghai 200241, China

^*Correspondence: Fang Chang (fangchang@fudan.edu.cn)

Received:2024-09-24 Accepted:2025-07-30 Online:2025-08-28
Supported by:
This work was supported by the grant from the Ministry of Science and Technology, People's Republic of China (2023YFA0913500), grants from the National Natural Science Foundation of China (32270347, 31870294, and 32470348) to F.C, and the China Postdoctoral Science Foundation (2023M740705) to Y.Y.

Abstract

Abstract: The pollen exine serves as a protective barrier and signaling interface essential for male fertility in flowering plants. Its precise patterning depends on coordinated interactions between microspores and tapetal cells. While the CLAVATA3/EMBRYO SURROUNDING REGION-related 19 (CLE19) peptide has been identified as a microspore-derived “brake” that restricts tapetal activity to maintain exine developmental homeostasis, how CLE19 integrates with hormonal signaling pathways remains poorly understood. Here, we demonstrate that CLE19 attenuates brassinosteroid (BR) signaling output by engaging a defined BSL–BIN2–BES1 signaling cascade. Through quantitative phosphoproteomic analysis, we identified that CLE19 affects the phosphorylation of multiple BR signaling components, including BSL-type phosphatases BSL1/2/3, the GSK3-like kinase BIN2, and the transcription factor BES1. We show that CLE19 is perceived by its receptor PXL1, which directly interacts with BSL-type phosphatases to activate the GSK3-like kinase BIN2, leading to phosphorylation of BES1 at serine residues S219 and S223. Functional analyses using phospho-dead and phospho-mimic BES1 variants confirm that CLE19-dependent phosphorylation controls BES1 nuclear export and degradation, ultimately suppressing BR-responsive transcriptional outputs required for pollen exine patterning. Together, our findings define a peptide–hormone signaling axis that regulates transcription factor activity through post-translational regulation, providing mechanistic insight into how developmental robustness is maintained via intercellular signal integration in plant reproduction.

Key words: Arabidopsis, BES1, brassinosteroid signaling, BSLs, CLE19, nuclear export, peptide–hormone crosstalk, phospho-proteomics, pollen exine, tapetum

Shuangshuang Wang, Shiting Zhang, Ying Yu, Jianzheng Wang, Jingya Wang, Mengyu Li, Jianan Lu, Juanying Ye, Hanji Li, Yeqiao Liu, Yuhan Zhao, Wen Song, Juan Dong, Jia Li, Chunming Liu, Hong Ma, Fang Chang. CLE19 suppresses brassinosteroid signaling output via the BSL-BIN2 module to maintain BES1 activity and pollen exine patterning in Arabidopsis[J]. J Integr Plant Biol., DOI: 10.1111/jipb.70024.

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CLE19 suppresses brassinosteroid signaling output via the BSL-BIN2 module to maintain BES1 activity and pollen exine patterning in Arabidopsis

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