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Published on: October 14, 2022
Molecular mechanism of DNA recognition by the rice zinc finger transcription factor Bsr-d1
Lifeng Ji1, Jie Jiang1, Yiling Qin1
1State Key Laboratory of Maize Bio-breeding, Joint International Research Laboratory of Crop Molecular Breeding, Ministry of Agriculture Key Laboratory for Crop Pest Monitoring and Green Control, College of Plant Protection, China Agricultural University, Beijing, China.
Abstract:
The rice transcription factor bsr-d1, a zinc finger protein, is a target for Magnaporthe oryzae pathogenesis, activating the expression of peroxidase genes and suppressing H2O2 generated in rice immunity. However, the specific recognition mechanism of Bsr-d1 and its target DNA is unknown. Here, with the structural prediction using AlphaFold3, we demonstrate that Bsr-d1 contains two Cys2-His2 zinc finger domains (ZF1:73-97 and ZF2:135-159) connected via an extended loop. Gel filtration and bio-layer interferometry (BLI) assays revealed that ZF1, but not ZF2, specifically binds the GTACT cis-element in peroxidase gene promoters. Guided by these findings, we propose an AlphaFold3-derived interaction model of the DNA/bsr-d1 in which residues Y85 and Q86, along with three positively charged residues (K92, R96, K97), are positioned close proximity to DNA binding sites. Functional validation showed that alanine substitutions at Y85, Q86, K92, R96 or K97 significantly disrupted DNA binding, while mutations in the GTACT motif of promoter region abolished Bsr-d1/promoter interaction. Collectively, our findings elucidate the structural basis of zinc finger-DNA recognition in plant immunity and establish a framework for engineering Bsr-d1 to confer rice broad-spectrum resistance.
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