The plant hormone jasmonate (JA) plays crucial roles in regulating plant responses to herbivorous insects and microbial pathogens and is an important regulator of plant growth and development1–7. Key mediators of JA signaling include MYC transcription factors, which are repressed by JAZ transcriptional repressors at the resting state. In the presence of active JA, JAZ proteins function as JA co-receptors by forming a hormone-dependent complex with COI1, the F-box subunit of an SCF-type ubiquitin E3 ligase8–11. The hormone-dependent formation of the COI1–JAZ co-receptor complex leads to ubiquitination and proteasome-dependent degradation of JAZ repressors and release of MYC proteins from transcriptional repression3,10,12. The mechanism by which JAZ proteins repress MYC transcription factors and how JAZ proteins switch between the repressor function in the absence of hormone and the co-receptor function in the presence of hormone remain enigmatic. Here we show that Arabidopsis MYC3 undergoes pronounced conformational changes when bound to the conserved Jas motif of the JAZ9 repressor. The Jas motif, previously shown to bind to hormone as a partially unwound helix, forms a complete α-helix that displaces the N-terminal helix of MYC3 and becomes an integral part of the MYC N-terminal fold. In this position, the Jas helix competitively inhibits MYC3 interaction with the MED25 subunit of the transcriptional Mediator complex. Our study elucidates a novel molecular switch mechanism that governs the repression and activation of a major plant hormone pathway.

Strigolactones (SLs) are endogenous hormones and exuded signaling molecules in plant responses to low levels of mineral nutrients. Key mediators of the SL signaling pathway in rice include the α/β-fold hydrolase DWARF 14 (D14) and the F-box component DWARF 3 (D3) of the ubiquitin ligase SCF D3 that mediate ligand-dependent degradation of downstream signaling repressors. One perplexing feature is that D14 not only functions as the SL receptor but is also an active enzyme that slowly hydrolyzes diverse natural and synthetic SLs including GR24, preventing the crystallization of a binary complex of D14 with an intact SL as well as the ternary D14/SL/D3 complex. Here we overcome these barriers to derive a structural model of D14 bound to intact GR24 and identify the interface that is required for GR24-mediated D14-D3 interaction. The mode of GR24-mediated signaling, including ligand recognition, hydrolysis by D14, and ligand-mediated D14-D3 interaction, is conserved in structurally diverse SLs. More importantly, D14 is destabilized upon the binding of ligands and D3, thus revealing an unusual mechanism of SL recognition and signaling, in which the hormone, the receptor, and the downstream effectors are systematically destabilized during the signal transduction process.

Membrane proteins impose enormous challenges to cellular protein homeostasis during their post-translational targeting, and require chaperones to keep them soluble and translocation-competent. Here we show that a novel targeting factor in the chloroplast Signal Recognition Particle (cpSRP), cpSRP43, is a highly specific molecular chaperone that efficiently reverses the aggregation of its substrate proteins. In contrast to AAA+-chaperones, cpSRP43 utilizes specific binding interactions with its substrate to mediate its disaggregase activity. This ‘disaggregase’ capability can allow targeting machineries to more effectively capture their protein substrates, and emphasizes a close connection between protein folding and trafficking processes. Moreover, cpSRP43 provides the first example of an ATP-independent disaggregase, and demonstrates that efficient reversal of protein aggregation can be attained by specific binding interactions between a chaperone and its substrate.

Magnetic random access memories based on the spin transfer torque phenomenon (STT-MRAMs) have become one of the leading candidates for next generation memory applications. Among the many attractive features of this technology are its potential for high speed and endurance, read signal margin, low power consumption, scalability, and non-volatility. In this paper, we discuss our recent results on perpendicular STT-MRAM stack designs that show STT efficiency higher than 5 kBT/μA, energy barriers higher than 100 kBT at room temperature for sub-40 nm diameter devices, and tunnel magnetoresistance higher than 150%. We use both single device data and results from 8 Mb array to demonstrate data retention sufficient for automotive applications. Moreover, we also demonstrate for the first time thermal stability up to 400 °C exceeding the requirement of Si CMOS back-end processing, thus opening the realm of non-volatile embedded memory to STT-MRAM technology.

We have characterized the ligand-enhanced phosphorylation of the CXC chemokine receptor-2 (CXCR2) in a series of clonal 3ASubE cell lines expressing receptors truncated or mutated in the carboxyl-terminal domain. Truncation of CXCR2 by substitution of a stop codon for Ser-342 (342T) or Ser-331 (331T) results in total loss of melanoma growth stimulatory activity/growth-related protein (MGSA/GRO)-enhanced receptor phosphorylation, which cannot be explained based upon altered ligand binding affinity or receptor number. 3ASubE cells expressing 342T or CXCR2 with mutation of Ser-342, -346, -347, and -348 to alanine (4A) exhibit strong mobilization of Ca 2؉ in response to ligand (interleukin-8 or MGSA/GRO), with a recovery phase significantly slower than that of cells expressing wild type (WT) CXCR2. In contrast to the WT CXCR2, which is 93% desensitized by 20 nM ligand, the 331T, 342T, and 4A CXCR2 mutants do not undergo significant ligand-induced desensitization, and respond to a second ligand challenge by mobilizing Ca 2؉ . The 3ASubE cells expressing CXCR2 with mutation of Ser-346, -347, and -348 to alanine, or with mutation of only one serine in this domain, continue to be phosphorylated in response to ligand and are 60 -70% desensitized following the initial ligand challenge. WT CXCR2 phosphorylation and desensitization occur in <1 min, while receptor sequestration is a much later event (30 -60 min). However, mutant receptors that are neither phosphorylated nor desensitized in response to ligand are <10% sequestered 60 min following ligand challenge. These data demonstrate for the first time that ligand binding to CXCR2 results in receptor phosphorylation, desensitization, and sequestration and that serine residues 342 and 346 -348 participate in the desensitization and sequestration processes.The CXC chemokine, 1 IL-8, specifically binds the CXC chemokine receptor, CXCR1, while a second receptor, CXCR2, is shared by IL-8, MGSA/GRO, and several other CXC chemokines (1). In human neutrophils, a 10-min treatment with IL-8 induces the internalization of greater than 90% of the CXC chemokine receptors, CXCR1 and CXCR2 (2). Low MGSA/GRO concentrations (0.2 nM) down-regulated 50% of CXCR2 expressed on neutrophils, while 7-13-fold higher IL-8 concentrations are required to down-regulate 50% of CXCR1 (3). Cell surface expression of CXCR1 is 100% restored after 1.5 h, while only 40% of the initial CXCR2 expression can be detected after 3 h of incubation following ligand treatment (3). We have previously demonstrated in both 3ASubE and 293 stable transfectants expressing CXCR2 that ligand binding results in the phosphorylation of CXCR2 on serine residues (4, 5). Within 1 min after ligand binding, MGSA/GRO stimulates the phosphorylation of CXCR2, and after prolonged treatment with MGSA/ GRO ϳ40% of the receptors are degraded (4). Phorbol ester treatment (100 nM for 2 h) of 3ASubE cells expressing CXCR2 also results in serine phosphorylation, down-regulation, and degradation of the receptor (5). These data suggest that phos...