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Nat Commun. 2019 Jul 19;10(1):3252. doi: 10.1038/s41467-019-11202-z.

TurboID-based proximity labeling reveals that UBR7 is a regulator of N NLR immune receptor-mediated immunity.

Author information

1
Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, 95616, USA. cauzhangyl@cau.edu.cn.
2
State Key Laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. cauzhangyl@cau.edu.cn.
3
Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA, 50011, USA.
4
Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, 95616, USA.
5
Departments of Genetics, Stanford University, Stanford, CA, 94305, USA.
6
Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
7
Department of Biology, Stanford University, Stanford, CA, 94305, USA.
8
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
9
Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA.
10
Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA, 50011, USA. jwalley@iastate.edu.
11
Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, 95616, USA. spdineshkumar@ucdavis.edu.

Abstract

Nucleotide-binding leucine-rich repeat (NLR) immune receptors play a critical role in defence against pathogens in plants and animals. However, we know very little about NLR-interacting proteins and the mechanisms that regulate NLR levels. Here, we used proximity labeling (PL) to identify the proteome proximal to N, which is an NLR that confers resistance to Tobacco mosaic virus (TMV). Evaluation of different PL methods indicated that TurboID-based PL provides more efficient levels of biotinylation than BioID and BioID2 in plants. TurboID-based PL of N followed by quantitative proteomic analysis and genetic screening revealed multiple regulators of N-mediated immunity. Interestingly, a putative E3 ubiquitin ligase, UBR7, directly interacts with the TIR domain of N. UBR7 downregulation leads to an increased amount of N protein and enhanced TMV resistance. TMV-p50 effector disrupts the N-UBR7 interaction and relieves negative regulation of N. These findings demonstrate the utility of TurboID-based PL in plants and the N-interacting proteins we identified enhance our understanding of the mechanisms underlying NLR regulation.

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