Abstract
Resistance to drought stress is fundamental to plant survival and development. Abscisic acid (ABA) is one of the major hormones involved in different types of abiotic and biotic stress responses. ABA intracellular signaling has been extensively explored in Arabidopsis thaliana and occurs via a phosphorylation cascade mediated by three related protein kinases, denominated SnRK2s (SNF1-related protein kinases). However, the role of ABA signaling and the biochemistry of SnRK2 in crop plants remains underexplored. Considering the importance of the ABA hormone in abiotic stress tolerance, here we investigated the regulatory mechanism of sugarcane SnRK2s—known as stress/ABA-activated protein kinases (SAPKs). The crystal structure of ScSAPK10 revealed the characteristic SnRK2 family architecture, in which the regulatory SnRK2 box interacts with the kinase domain αC helix. To study sugarcane SnRK2 regulation, we produced a series of mutants for the protein regulatory domains SnRK2 box and ABA box. Mutations in ScSAPK8 SnRK2 box aimed at perturbing its interaction with the protein kinase domain reduced protein kinase activity in vitro. On the other hand, mutations to ScSAPK ABA box did not impact protein kinase activity but did alter the protein autophosphorylation pattern. Taken together, our results demonstrate that both SnRK2 and ABA boxes might play a role in sugarcane SnRK2 function.
Highlights
The phytohormone abscisic acid (ABA) is a central regulator of plant responses to abiotic stress
Three Abscisic acid (ABA)-responsive SnRK2s were identified within the sugarcane genome (S. spontaneum × S. officinarum hybrid cultivar) using homologous protein sequences from A. thaliana and Sorghum bicolor
Phylogenetic analysis revealed that sugarcane SnRK2s present higher homology to Z. mays SAPK8 compared to A. thaliana proteins (Supplementary Figure S1)
Summary
The phytohormone abscisic acid (ABA) is a central regulator of plant responses to abiotic stress. ABA triggers protective plant responses leading to stomatal closure, seed dormancy, inhibition of growth, and germination (Mustilli, 2002; Yoshida et al, 2002; Yoshida et al, 2006; Fujii et al, 2007; Fujii and Zhu, 2009). Members of the SnRK2 sub-family of serine-threonine kinases (SnRK2.2/2.3/2.6 in Arabidopsis) are the positive regulators of ABA signaling and activate downstream transcription factors, leading to the expression of stress-responsive genes (Fujii and Zhu, 2009; Fujii et al, 2009; Fujita et al, 2009; Nakashima et al, 2009). ABA-responsive kinases from both mono- and eudicotyledons are expected to have a conserved modular architecture and to be involved in environmental sensing and stress response (Kulik et al, 2011)
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