AP2/ERFs contain a highly conserved DNA-binding domain consisting of approximately 60 amino acids (Nakano et al. Additionally, an AP2-LIKE ERE BINDING FACTOR (ERF), also known as ERE BINDING PROTEIN (EREBP), was first isolated as the GCC box-binding protein from tobacco (Ohme-Takagi and Shinshi 1995). APETALA2 was first isolated from the floral development-related proteins in Arabidopsis (Jofuku et al. APETALA2/ethylene response factors (AP2/ERFs) are a superfamily of plant-exclusive transcription factors involved in the ethylene-inducible response (Gutterson and Reuber 2004 Mizoi et al. Such a group of transcription factors in plants is activated by ethylene signaling under biotic or abiotic stresses. A cis-acting element GCC box motif that constitutes the conserved sequence AGCCGCC is widely present in the promoter region of ethylene-inducible genes and has been suggested as a core sequence of the ethylene-responsive element (ERE) for ethylene signaling in plants (Buttner and Singh 1997 Hao et al. These stresses can induce the biosynthesis of ethylene, a gaseous plant hormone confirmed as a mediator of plant stress responses (Ecker 1995). 2019), such as herbivore damage, pathogenic infection, UV irradiation, temperature variation, drought, and high salt content (Bechtold and Field 2018 Kazan 2015 Penninckx et al. To rapidly adapt to environmental change, numerous genes are regulated in response to biotic and abiotic stresses (Xie et al. Plants are exposed to natural environments that may negatively affect their growth and development. Our study establishes the structural basis of the AtERF96 transcription factor in complex with the GCC box, as well as the DNA binding mechanisms of the N-terminal α-helix and AP2/ERF domain. We also demonstrated the structure of AtERF96 EDLL motif, a unique conserved motif in the group IX of AP2/ERF family, might involve in the transactivation of defense-related genes. In addition to the AP2/ERF domain, an N-terminal α-helix of AtERF96 participates in DNA interaction in the flanking region.
We analyzed the binding residues of the conserved AP2/ERF domain in the DNA recognition sequence. In this report, we determined the crystal structure of AtERF96 in complex with a GCC box at atomic resolution. A previous study showed only the NMR structure of the AP2/ERF domain of AtERF100 in complex with a GCC box DNA motif. Ethylene-responsive element binding protein, a member of the APETALA2/ethylene response factor (AP2/ERF) superfamily, is a transcription factor that regulates stress-responsive genes by recognizing a specific cis-acting element of target DNA. The phytohormone ethylene is widely involved in many developmental processes and is a crucial regulator of defense responses against biotic and abiotic stresses in plants.
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