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- W2889966424 abstract "B3 transcription factors constitute a large plant‐specific protein superfamily, which plays a central role in plant life. Family members are characterized by the presence of B3 DNA ‐binding domains ( DBD s). To date, only a few B3 DBD s were structurally characterized; therefore, the DNA recognition mechanism of other family members remains to be elucidated. Here, we analyze DNA recognition mechanism of two structurally uncharacterized B3 transcription factors, ABI 3 and NGA 1. Guided by the structure of the DNA ‐bound B3 domain of Arabidopsis transcriptional repressor VAL 1, we have performed mutational analysis of the ABI 3 B3 domain. We demonstrate that both VAL 1‐B3 and ABI 3‐B3 recognize the Sph/ RY DNA sequence 5′‐ TGCATG ‐3′ via a conserved set of base‐specific contacts. We have also solved a 1.8 Å apo‐structure of NGA 1‐B3, DBD of Arabidopsis transcription factor NGA 1. We show that NGA 1‐B3, like the structurally related RAV 1‐B3 domain, is specific for the 5′‐ CACCTG ‐3′ DNA sequence, albeit tolerates single base pair substitutions at the 5′‐terminal half of the recognition site. Employing distance‐dependent fluorophore quenching, we show that NGA 1‐B3 binds the asymmetric recognition site in a defined orientation, with the ‘N‐arm’ and ‘C‐arm’ structural elements interacting with the 5′‐ and 3′‐terminal nucleotides of the 5′‐ CACCTG ‐3′ sequence, respectively. Mutational analysis guided by the model of DNA ‐bound NGA 1‐B3 helped us identify NGA 1‐B3 residues involved in base‐specific and DNA backbone contacts, providing new insights into the mechanism of DNA recognition by plant B3 domains of RAV and REM families. Databases RCSB Protein Data Bank, accession number 5OS9 ." @default.
- W2889966424 created "2018-09-27" @default.
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- W2889966424 date "2018-09-21" @default.
- W2889966424 modified "2023-09-26" @default.
- W2889966424 title "<scp>DNA</scp> recognition by <i>Arabidopsis</i> transcription factors <scp>ABI</scp> 3 and <scp>NGA</scp> 1" @default.
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- W2889966424 doi "https://doi.org/10.1111/febs.14649" @default.
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