Single-Molecule and Vesicle Trafficking Analysis of Ubiquitination Involved in the Activity of Ammonium Transporter AMT1;3 in Arbidopsis under High Ammonium Stress

Ran Zhao; Yangyang Cao; Yanrui Ge; Jing Xu; Ruofan Li; Mei Yang; Yingying Chen; Dingjie Wu; Jianwei Xiao; Ruili Li

doi:10.3390/cells11223651

Single-Molecule and Vesicle Trafficking Analysis of Ubiquitination Involved in the Activity of Ammonium Transporter AMT1;3 in Arbidopsis under High Ammonium Stress

Cells. 2022 Nov 17;11(22):3651. doi: 10.3390/cells11223651.

Authors

Ran Zhao^{1

2}, Yangyang Cao^{1

2

3}, Yanrui Ge^{1

2}, Jing Xu^{1

2}, Ruofan Li^{1

2}, Mei Yang^{1

2}, Yingying Chen^{1

2}, Dingjie Wu^{1

2}, Jianwei Xiao^{1

2}, Ruili Li^{1

2}

Affiliations

¹ National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China.
² Institute of Tree Development and Genome Editing, Beijing Forestry University, Beijing 100083, China.
³ Shijiazhuang Zhonghua Avenue Primary School, Shijiazhuang 050000, China.

Abstract

Plants absorb nitrogen from the soil using ammonium transporters (AMTs). Plants can precisely regulate AMT1;3 levels using sophisticated regulatory systems, ensuring adequate nitrogen uptake without hazardous ammonium production. Here, we demonstrated that ubiquitylation can contribute to AMT1;3 degradation under high ammonium stress. Using the ubiquitin site mutant AMT1;3_K75R,K233R-EGFP, we demonstrated that the loss of ubiquitination affects the dynamic characteristics of AMT1;3 proteins on the plasma membrane and markedly inhibits the endocytosis of AMT1;3 proteins under high ammonium stress. AMT1;3_K75R,K233R-EGFP plants also showed inhibition of protein degradation that targets the vesicular pathway after being exposed to high levels of ammonium. Our findings showed that the dynamic properties, endocytosis, and vesicle trafficking pathways of AMT1;3 proteins are altered in AMT1;3_K75R,K233R-EGFP under high ammonium conditions.

Keywords: 3; AMT1; dynamics; endocytosis; ubiquitylation; vesicle trafficking.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ammonium Compounds* / metabolism
Ammonium Compounds* / pharmacology
Arabidopsis* / metabolism
Cation Transport Proteins* / genetics
Cation Transport Proteins* / metabolism
Nitrogen / metabolism
Plant Proteins / genetics
Plant Proteins / metabolism
Ubiquitination

Substances

Ammonium Compounds
Cation Transport Proteins
Plant Proteins
Nitrogen

Grants and funding

The research reported in this publication was supported by the National Natural Science Foundation of China (32270368, 31970182 and 31761133009), the Fundamental Research Funds for the Central Universities (2021ZY57) and the National Training Program of Innovation and Entrepreneurship for Undergraduates (202110022019, 202110022069).