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Proteins, Expression, Isolation and Analysis> | … For the pull-down assays of SPL9 and ABI5, the fusion proteins were incubated with glutathione resin (GenScript) overnight in column buffer (20 mM Tris–HCl [pH 7.5], 200 mM NaCl, 1 mM PMSF, 1 mM DTT and 1× protease inhibitor (Roche 4693132001)]. For the pull-down … | Get A Quote |
The miR156-targeted SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors play key roles in regulating plant development, but little is known about their function in abscisic acid (ABA) signaling. Here, we report that the miR156-targeted SPLs enhance ABA responses and contribute to the inhibition of pre-harvest sprouting. We find that SPL9 directly activates the expression of ABA responsive genes through binding to their promoters. SPL9 was further shown to physically interact with ABSCISIC ACID INSENSITIVE 5 (ABI5), a master transcription factor in ABA signaling, thus promoting its association with the promoters of ABA responsive genes. Furthermore, we reveal that the protein kinases SnRK2s intera... More