Abscisic acid (ABA) is a ubiquitous hormone that regulates plant growth, development, and responses to environmental stresses. Its action is mediated by the PYR/PYL/RCAR family of START proteins, but it remains unclear how these receptors bind ABA and in turn, how hormone binding leads to inhibition of the downstream type 2C protein phosphatase (PP2C) effectors. Here we report crystal structures of apo and ABA-bound receptors as well as a ternary PYL2-ABA-PP2C complex. The apo receptors contain an open ligand-binding pocket flanked by a gate that closes in response to ABA via conformational changes in two highly conserved β-loops that serve as a gate and latch. Moreover, ABA-induced closure of the gate creates a surface that enables the receptor to dock into and competitively inhibit the PP2C active site. A conserved tryptophan in the PP2C inserts directly between the gate and latch, which functions to further lock the receptor in a closed conformation. Together, our results identify a conserved gate-latch-lock mechanism underlying ABA signaling.
SummaryA series of papers in the last year reported major advances in our understanding of ABA signaling: the identification of soluble ABA receptors, the elucidation of a core ABA signaling pathway and structural insights into the mechanism of ABA perception and signaling. Here we summarize these advances, which have shown in atomic resolution that the ABA receptors PYR1, PYL1 and PYL2 function as allosteric switches that inhibit type 2C protein phosphatases (PP2Cs) in response to ABA. These receptors function at the apex of a core signaling pathway that regulates ABA responses by controlling SnRK2 kinase activity and the phosphorylation of downstream target proteins such as ABFs, which control nuclear responses, and the ion channel SLAC1, which mediates electrophysiological responses to ABA.
In this investigation, a confirmed case in a household contact was defined as having received a positive SARS-CoV-2 nucleic acid amplification test result or antigen test result ≤14 days after the index date (date of the index patient's symptom onset or positive SARS-CoV-2 nucleic acid amplification test result or antigen test result), and a probable case in a household contact was defined as the presence of COVID-19-compatible symptoms during the same 14-day period but without a positive SARS-CoV-2 test confirmation. Persons without symptoms and who did not have a positive SARS-CoV-2 test result were not considered to have a case of COVID-19. Analysis of AR among household contacts excluded eight persons with unknown case status (persons for whom it was not known whether COVID-19-compatible symptoms were present and whether SARS-CoV-2 testing had occurred [or if testing occurred, the results were unknown]).
Changing environmental conditions and lessening fresh water supplies have sparked intense interest in understanding and manipulating abscisic acid signaling, which controls adaptive responses to drought and other abiotic stressors. We recently discovered a selective ABA agonist, pyrabactin, and used it to discover its primary target PYR1, the founding member of the PYR/PYL family of soluble ABA receptors. To understand pyrabactin's selectivity we have taken a combined structural, chemical and genetic approach. We show that subtle differences between receptor binding pockets control ligand orientation between productive and non-productive modes. Non-productive binding occurs without gate closure and prevents receptor activation. Observations in solution show that these orientations are in rapid equilibrium that can be shifted by mutations to control maximal agonist activity. Our results provide a robust framework for the design of new agonists and reveal a new mechanism for agonist selectivity.
The chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) directs leukocyte migration, stem cell homing, and cancer metastasis through activation of CXCR4, which is also a coreceptor for T-tropic HIV-1. Recently, SDF-1 was shown to play a protective role after myocardial infarction, and the protein is a candidate for development of new anti-ischemic compounds. SDF-1 is monomeric at nanomolar concentrations but binding partners promote self-association at higher concentrations to form a typical CXC chemokine homodimer. Two NMR structures have been reported for the SDF-1 monomer, but only one matches the conformation observed in a series of dimeric crystal structures. In the other model, the C-terminal helix is tilted at an angle incompatible with SDF-1 dimerization. Using a rat heart explant model for ischemia/ reperfusion injury, we found that dimeric SDF-1 exerts no cardioprotective effect, suggesting that the active species is monomeric. To resolve the discrepancy between existing models, we solved the NMR structure of the SDF-1 monomer in different solution conditions. Irrespective of pH and buffer composition, the C-terminal helix remains tilted at an angle with no evidence for the perpendicular arrangement. Furthermore, we find that phospholipid bicelles promote dimerization that necessarily shifts the helix to the perpendicular orientation, yielding dipolar couplings that are incompatible with the NOE distance constraints. We conclude that interactions with the alignment medium biased the previous structure, masking flexibility in the helix position that may be essential for the distinct functional properties of the SDF-1 monomer.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.