Onses are thought to be insufficient to suppress the ABA hypersensitivity of agb1 fully. It’s also fascinating that BZR1 overexpression as an alternative to bzr1-1 overexpression alleviated the ABA effects (Fig. 3A). BZR1 and bzr1-1 have already been suggested to differ in their protein stabilities (He et al., 2002; Wang et al., 2002; Tang et al., 2011), phosphorylation states (He et al., 2002; Tang et al., 2011), and subcellular localizations (Ryu et al., 2007; this study, Supplementary Fig. S12 at JXB on the net). In addition, bzr1-1 features a greater affinity for a subunit of PP2A than does BZR1 (Tang et al., 2011). BZR1 and bzr1-1 may possibly have unique affinities for other interaction partners that affect the expression of BZR1 target genes. These differences may well cause the various effects of BZR1 and bzr1-1 around the gene expression involved in the ABA responses.Canagliflozin Additional studies are needed to elucidate the molecular mechanisms underlying BZR1mediated ABA signalling. AGB1 was shown to interact with BIN2 in vitro (Fig. 4B). Nevertheless, AGB1 didn’t impact the kinase activity of BIN2 (Figs 4B, 5B) and was not phoshorylated by BIN2 (Fig. 4B). Hence the physiological relevance from the interaction among AGB1 and BIN2 is unclear. One possibility is the fact that AGB1 regulates the phosphorylation of BIN2 substrates besides BZR1 and BZR2. ARF2 and YODA are two examples of BIN2 substrates apart from BZR1. ARF2 can be a transcription factor regulating auxin-responsive gene expression (Vert et al., 2008). YODA is really a mitogen-activated protein kinase kinase kinase (MAPKKK) regulating stomatal improvement, and, interestingly, YODA has been suggested to be below the handle of ER signalling as well as BIN2-dependent signalling (Kim et al., 2012). AGB1 is also involved in auxin signalling in Arabidopsis (Ullah et al., 2003), and Pisum sativum G interacts with a MAPK (Bhardwaj et al., 2011). Thus, it will be intriguing to examine no matter if AGB1 regulates the BIN2-dependent phosphorylation of ARF2 and YODA.Supplementary dataSupplementary data are readily available at JXB on-line. Figure S1. Phenotypes of agb1 grown in diverse concentrations of BRZ. Figure S2. Absolute hypocotyl lengths of agb1 grown inside the presence of BRZ.Xanthohumol Figure S3.PMID:24013184 BR-induced hypocotyl elongation in agb1. Figure S4. Semi-quantitative RT CR evaluation of BZR1GFP and bzr1-1-GFP expression. Figure S5. Absolute hypocotyl lengths of bzr1-1 FPoverexpressing plants grown in the presence of BRZ. Figure S6. Expression levels of BZR1-GFP and bzr1-1GFP in transgenic plants. Figure S7. BZR1 expression levels in BZR1-GFPoverexpressing lines and their responses to ABA. Figure S8. Expression of His-AGB1 and GST IN2 in E. coli. Figure S9. In vitro GST pull-down assay. Figure S10. BiFC amongst AGB1 and BIN2. Figure S11. Effects of AGB1 on the interaction amongst BIN2 and BZR1 within a Y3H method. Figure S12. Subcellular localizations of BZR1 FP in agb1. Table S1. Primer pairs made use of for RT CR analyses.AcknowledgementsThis operate was supported by a Grant-in-aid for Scientific Investigation (21380002) to TT and (22144) to DT. We are grateful for the ABRC for giving the Arabidopsis mutant seeds. An Arabidopsis full-length cDNA clone (RAFL0420-E20) was developed by the plant genome project of RIKEN Genomic Sciences Center, and supplied by RIKEN BRC which can be participating inside the National Bio-Resource Project from the MEXT, Japan.3222 | Tsugama et al.
FEBS Open Bio 3 (2013) 352journal homepage: www.elsevier/locate/febsopenbioMurine homologue with the human.
