RESUMEN
With the advantages of high energy density, high accuracy, and fast response, smart material-driven electro-hydrostatic actuators (SMEHAs) have attracted significant attention in recent years. However, the low flow rate of SMEHAs constrains their application. One potential solution to enhance the flow rate is to increase the number of smart material-actuated pumps. In view of this, this paper proposes a new configuration of an electro-hydrostatic actuator equipped with four magnetostrictive-actuated pumps (FMEHA) to achieve a large flow rate. The mathematical model of the FMEHA is established to investigate the driving phase matching between pumps and the active flow distribution valve. The physical prototype of FMEHA is fabricated. Simulations and experiments are conducted to assess its performance under various driving parameters, including the number of pumps, driving phase, frequency, and amplitude. The optimal driving parameters for the FMEHA are determined based on the results obtained. Experimental findings demonstrate that with a driving phase of 340°, a frequency of 250 Hz, and an amplitude of 20 A, the FMEHA achieves a maximum flow rate of 6.2 l/min.
RESUMEN
Phosphate, a key plant nutrient, is perceived through inositol polyphosphates (InsPs) by SPX domain-containing proteins. SPX1 an inhibit the PHR2 transcription factor to maintain Pi homeostasis. How SPX1 recognizes an InsP molecule and represses transcription activation by PHR2 remains unclear. Here we show that, upon binding InsP6, SPX1 can disrupt PHR2 dimers and form a 1:1 SPX1-PHR2 complex. The complex structure reveals that SPX1 helix α1 can impose a steric hindrance when interacting with the PHR2 dimer. By stabilizing helix α1, InsP6 allosterically decouples the PHR2 dimer and stabilizes the SPX1-PHR2 interaction. In doing so, InsP6 further allows SPX1 to engage with the PHR2 MYB domain and sterically block its interaction with DNA. Taken together, our results suggest that, upon sensing the surrogate signals of phosphate, SPX1 inhibits PHR2 via a dual mechanism that attenuates dimerization and DNA binding activities of PHR2.
Asunto(s)
Proteínas de Arabidopsis/química , Arabidopsis/metabolismo , ADN de Plantas/química , Fosfatos de Inositol/metabolismo , Proteínas Nucleares/química , Oryza/metabolismo , Arabidopsis/química , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Sitios de Unión , Clonación Molecular , Cristalografía por Rayos X , ADN de Plantas/genética , ADN de Plantas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Regulación de la Expresión Génica de las Plantas , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Fosfatos de Inositol/química , Modelos Moleculares , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Nutrientes/química , Nutrientes/metabolismo , Oryza/química , Oryza/genética , Plantas Modificadas Genéticamente , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Multimerización de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transducción de SeñalRESUMEN
The maintenance of cellular phosphate (Pi) homeostasis is of great importance in living organisms. The SPX domain-containing protein 1 (SPX1) proteins from both Arabidopsis and rice have been proposed to act as sensors of Pi status. The molecular signal indicating the cellular Pi status and regulating Pi homeostasis in plants, however, remains to be identified, as Pi itself does not bind to the SPX domain. Here, we report the identification of the inositol pyrophosphate InsP8 as a signaling molecule that regulates Pi homeostasis in Arabidopsis. Polyacrylamide gel electrophoresis profiling of InsPs revealed that InsP8 level positively correlates with cellular Pi concentration. We demonstrated that the homologs of diphosphoinositol pentakisphosphate kinase (PPIP5K), VIH1 and VIH2, function redundantly to synthesize InsP8, and that the vih1 vih2 double mutant overaccumulates Pi. SPX1 directly interacts with PHR1, the central regulator of Pi starvation responses, to inhibit its function under Pi-replete conditions. However, this interaction is compromised in the vih1 vih2 double mutant, resulting in the constitutive induction of Pi starvation-induced genes, indicating that plant cells cannot sense cellular Pi status without InsP8. Furthermore, we showed that InsP8 could directly bind to the SPX domain of SPX1 and is essential for the interaction between SPX1 and PHR1. Collectively, our study suggests that InsP8 is the intracellular Pi signaling molecule serving as the ligand of SPX1 for controlling Pi homeostasis in plants.