Random mutagenesis-based screening of the interface of phyllogen, a bacterial phyllody-inducing effector, for interaction with plant MADS-box proteins

Frontiers in Plant Science

Plant Science

Nozomu Iwabuchi¹  Kensaku Maejima¹  Juri Matsuyama¹  Yugo Kitazawa¹  Oki Matsumoto¹  Masato Suzuki¹  Yasuyuki Yamaji¹  Shigetou Namba¹  Hiroaki Koinuma¹  Kenro Oshima² 

[1] Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan;Faculty of Bioscience and Applied Chemistry, Hosei University, Tokyo, Japan;

关键词: MADS-box transcription factors;    phyllogen;    protein-protein interaction (PPI);    protein structure prediction (PSP);    random mutagenesis;   

DOI  :  10.3389/fpls.2023.1058059

 received in 2022-09-30, accepted in 2023-03-01,  发布年份 2023

来源: Frontiers

【 摘 要 】

To understand protein function deeply, it is important to identify how it interacts physically with its target. Phyllogen is a phyllody-inducing effector that interacts with the K domain of plant MADS-box transcription factors (MTFs), which is followed by proteasome-mediated degradation of the MTF. Although several amino acid residues of phyllogen have been identified as being responsible for the interaction, the exact interface of the interaction has not been elucidated. In this study, we comprehensively explored interface residues based on random mutagenesis using error-prone PCR. Two novel residues, at which mutations enhanced the affinity of phyllogen to MTF, were identified. These residues, and all other known interaction-involved residues, are clustered together at the surface of the protein structure of phyllogen, indicating that they constitute the interface of the interaction. Moreover, in silico structural prediction of the protein complex using ColabFold suggested that phyllogen interacts with the K domain of MTF via the putative interface. Our study facilitates an understanding of the interaction mechanisms between phyllogen and MTF.

【 授权许可】

Unknown   
Copyright © 2023 Kitazawa, Iwabuchi, Maejima, Matsumoto, Suzuki, Matsuyama, Koinuma, Oshima, Namba and Yamaji