RESUMEN
The S6 kinases (S6Ks) are known to be activated by the target of rapamycin through phosphorylation of their hydrophobic motif (HM). However, our previous research showed that the HM site of plant S6Ks is not phosphorylated and is not essential for their activity in yeast cells lacking Ypk3, an ortholog of mammalian S6K. Here, we demonstrate that the HM site of mammalian S6Ks is phosphorylated and is indispensable for their activity in yeast ypk3∆ cells. Furthermore, pseudo-phosphorylation at the HM site of plant S6Ks results in regaining of activity that is lost due to mutation in the conserved phosphorylation sites, namely the T-loop and Turn motif. These results indicate the activation mechanism of plant S6Ks is different from that of mammals.
Asunto(s)
Plantas/enzimología , Proteínas Quinasas S6 Ribosómicas/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Activación Enzimática , Humanos , Fosforilación , Unión Proteica , Proteínas Quinasas S6 Ribosómicas/química , Especificidad de la EspecieRESUMEN
INDETERMINATE DOMAIN (IDD) family proteins are plant-specific transcription factors. Some Arabidopsis IDD (AtIDD) proteins regulate the expression of SCARECROW (SCR) by interacting with GRAS family transcription factors SHORT-ROOT (SHR) and SCR, which are involved in root tissue formation. Some AtIDD proteins regulate genes involved in the synthesis (GA3ox1) or signaling (SCL3) of gibberellic acid (GA) by interacting with DELLA proteins, a subfamily of the GRAS family. We analyzed the DNA binding properties and protein-protein interactions of select AtIDD proteins. We also investigated the transcriptional activity of the combination of AtIDD and GRAS proteins (AtIDD proteins combined with SHR and SCR or with REPRESSOR of ga1-3 (RGA)) on the promoters of SCR,SCL3, and GA3ox1 by conducting a transient assay using Arabidopsis culture cells. Our results showed that the SCR promoter could be activated by the IDD and RGA complexes and that the SCL3 and GA3ox1 promoters could be activated by the IDD, SHR, and SCR complexes, indicating the possibility that these complexes regulate and consequently coordinate the expression of genes involved in GA synthesis (GA3ox1), GA signaling (SCL3), and root formation (SCR).
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proteínas Co-Represoras/genética , Oxigenasas de Función Mixta/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Unión al ADN/genética , Giberelinas/metabolismo , Familia de Multigenes/genética , Regiones Promotoras Genéticas/genética , Proteínas Represoras/genética , Factores de Transcripción/genéticaRESUMEN
Seed germination, one of the most important stages in a plant's life cycle, can be affected by abiotic stresses, such as salinity. The plant hormone abscisic acid (ABA) and high concentrations of glucose are also known to inhibit germination. In contrast, nitrate is known to stimulate germination in many plants. However, this stimulatory effect has not yet been investigated in the presence of inhibitory effects caused by abiotic stresses, ABA, and glucose. In this study, we show that nitrate can alleviate the inhibitory effects of sodium chloride (NaCl) or high concentrations of glucose on seed germination in Arabidopsis, while it was not able to promote germination that was inhibited by exogenous ABA and mannitol (an inducer of osmotic stress). An analysis of the gene expression involved in the regulation of germination showed that GA20ox1, encoding the gibberellin (GA) synthesis enzyme, SPATULA (SPT), encoding a bHLH transcription factor, and CYP707A2, encoding an ABA catabolic enzyme, were significantly upregulated by the addition of KNO3 in the presence of NaCl or glucose. Our results suggest the possibility that these genes are involved in the nitrate-mediated control of seed germination in the presence of NaCl or glucose.