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Mouse Double Minute 2 (MDM2) is a key negative regulator of the tumor suppressor protein p53. MDM2 overexpression occurs in many types of cancer and results in the suppression of WT p53. The 14-3-3 family of adaptor proteins are known to bind MDM2 and the 14-3-3σ isoform controls MDM2 cellular localization and stability to inhibit its activity. Therefore, small molecule stabilization of the 14-3-3σ/MDM2 protein-protein interaction (PPI) is a potential therapeutic strategy for the treatment of cancer. Here, we provide a detailed biophysical and structural characterization of the phosphorylation-dependent interaction between 14-3-3σ and peptides that mimic the 14-3-3 binding motifs within MDM2. The data show that di-phosphorylation of MDM2 at S166 and S186 is essential for high affinity 14-3-3 binding and that the binary complex formed involves one MDM2 di-phosphorylated peptide bound to a dimer of 14-3-3σ. However, the two phosphorylation sites do not simultaneously interact so as to bridge the 14-3-3 dimer in a 'multivalent' fashion. Instead, the two phosphorylated MDM2 motifs 'rock' between the two binding grooves of the dimer, which is unusual in the context of 14-3-3 proteins. In addition, we show that the 14-3-3σ-MDM2 interaction is amenable to small molecule stabilization. The natural product fusicoccin A forms a ternary complex with a 14-3-3σ dimer and an MDM2 di-phosphorylated peptide resulting in the stabilization of the 14-3-3σ/MDM2 PPI. This work serves as a proof-of-concept of the drugability of the 14-3-3/MDM2 PPI and paves the way toward the development of more selective and efficacious small molecule stabilizers., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

14-3-3 proteins are a family of regulatory hubs that function through a vast network of protein-protein interactions. Their dysfunction or dysregulation is implicated in a wide range of diseases, and thus they are attractive drug targets, especially for molecular glues that promote protein-protein interactions for therapeutic intervention. However, an incomplete understanding of the molecular mechanisms that underpin 14-3-3 function hampers progress in drug design and development. Biophysical methodologies are an essential element of the 14-3-3 analytical toolbox, but in many cases have not been fully exploited. Here, we present a contemporary review of the predominant biophysical techniques used to study 14-3-3 protein-protein interactions, with a focus on examples that address key questions and challenges in the 14-3-3 field., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Thurairajah, Hudson and Doveston.)

Interleukin-17 (IL-17)-producing γδ (γδ17) T cells are innate-like lymphocytes that contribute to protective anti-microbial responses but are also implicated in pathogenic inflammation at barrier sites. Understanding tissue-specific signals that regulate this subset is important to boost host defense mechanisms, but also to mitigate immunopathology. Here, we demonstrate that prostaglandin E 2 (PGE 2 ), a cyclooxygenase-dependent member of the eicosanoid family, directly enhances cytokine production by circulating and tissue-specific γδ17 T cells in vitro. Gain- and loss-of-function in vivo approaches further reveal that although provision of PGE 2 amplifies psoriasiform inflammation, ablation of host mPGES1-dependent PGE 2 synthesis is dispensable for cutaneous γδ17 T cell activation. By contrast, loss of endogenous PGE 2 production or depletion of the gut microbiota compromises intestinal γδ17 T cell responses and increases disease severity during experimental colitis. Together, our results demonstrate how a lipid mediator can synergize with tissue-specific signals to enhance innate lymphocyte production of IL-17 during barrier inflammation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Using an 8-year (2007-2014) data set from two different limb-viewing instruments, we evaluate the relative roles of vertically versus obliquely propagating gravity waves (GWs) as sources of GWs in the polar summer mesosphere. Obliquely propagating waves are of interest because they are presumed to be generated by the summer monsoons. In the high-latitude upper mesosphere, the correlation coefficient between the time series of ice water content (IWC) and GW amplitude is 0.48, indicating that the observed GWs enhance polar mesospheric clouds (PMCs). For vertically propagating waves, the correlation coefficient between IWC and stratospheric/lower mesospheric (20-70 km) GW amplitude at the same high latitudes becomes more negative with increasing altitude. This change in correlation from negative in the lower mesosphere to positive at PMC altitudes suggests the presence of another source of GWs. The positive correlation coefficient between the time series of IWC and GW amplitude from 0-50°N, 20-90 km shows a slanted structure suggesting oblique propagation. This slanted structure is more robust in some seasons compared to others, and this interannual variability may be due to the latitudinal gradient of the mesospheric easterly jet where steeper gradients allow for low-latitude tropospheric GWs to be refracted to the high-latitude mesosphere more efficiently. Gravity-Wave Regional or Global Ray Tracer (GROGRAT) ray tracing simulations show that more GWs propagate obliquely compared to vertically propagating waves that reach PMC altitudes. For obliquely propagating waves, GROGRAT simulations indicate that nonorographic tropospheric GWs with faster phase speed (>20 m/s) and longer horizontal wavelength (>400 km) have a higher probability of reaching the polar summer mesosphere.

Skin is a fundamental component of our host defense system that provides a dynamic physical and chemical barrier against pathogen invasion and environmental insults. Cutaneous barrier function is mediated by complex interactions between structural cells such as keratinocytes and diverse lineages of immune cells. In contrast to the protective role of these intercellular interactions, uncontrolled immune activation can lead to keratinocyte dysfunction and psoriasis, a chronic inflammatory disease affecting 2% of the global population. Despite some differences between human and murine skin, animal models of psoriasiform inflammation have greatly informed clinical approaches to disease. These studies have helped to identify the interleukin (IL)-23-IL-17 axis as a central cytokine network that drives disease. In addition, they have led to the recent description of long-lived, skin-resident innate lymphocyte and lymphoid cells that accumulate in psoriatic lesions. Although not completely defined, these populations have both overlapping and unique functions compared to antigen-restricted αβ T lymphocytes, the latter of which are well-known to contribute to disease pathogenesis. In this review, we describe the diversity of innate lymphocytes and lymphoid cells found in mammalian skin with a special focus on αβ T cells, Natural Killer T cells and Innate Lymphoid cells. In addition, we discuss the effector functions of these unique leukocyte subsets and how each may contribute to different stages of psoriasis. A more complete understanding of these cell types that bridge the innate and adaptive immune system will hopefully lead to more targeted therapies that mitigate or prevent disease progression., (Copyright © 2020 Polese, Zhang, Thurairajah and King.)

Interleukin-17-producing γδ T (γδ17) cells play a central role in protective and pathogenic immune responses. However, the tissue-specific mechanisms that control the activation of these innate lymphocytes are not known. Here, we demonstrate that CD109, a glycosylphosphatidylinositol (GPI)-anchored protein highly expressed by keratinocytes, is an important regulator of skin homeostasis and γδ17 cell activation. Genetic deletion of CD109 results in spontaneous epidermal hyperplasia, aberrant accumulation of dermal-derived γδ17 cells, and enhanced susceptibility to psoriasiform inflammation. In this context, γδ17 activation requires interleukin (IL)-23 signals and is reversed by transient depletion of the skin microbiota. Mechanistically, CD109 restrains γδ17 cell activation in a cell-extrinsic manner by fortifying skin barrier integrity. Collectively, our data provide insight into the regulation of the skin IL-23/IL-17 immune axis and how homeostasis is maintained at this important barrier site., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Measurements of turbulence and waves were made as part of the Mesosphere-Lower Thermosphere Turbulence Experiment (MTeX) on the night of 25-26 January 2015 at Poker Flat Research Range, Chatanika, Alaska (65°N, 147°W). Rocket-borne ionization gauge measurements revealed turbulence in the 70- to 88-km altitude region with energy dissipation rates between 0.1 and 24 mW/kg with an average value of 2.6 mW/kg. The eddy diffusion coefficient varied between 0.3 and 134 m 2 /s with an average value of 10 m 2 /s. Turbulence was detected around mesospheric inversion layers (MILs) in both the topside and bottomside of the MILs. These low levels of turbulence were measured after a minor sudden stratospheric warming when the circulation continued to be disturbed by planetary waves and winds remained weak in the stratosphere and mesosphere. Ground-based lidar measurements characterized the ensemble of inertia-gravity waves and monochromatic gravity waves. The ensemble of inertia-gravity waves had a specific potential energy of 0.8 J/kg over the 40- to 50-km altitude region, one of the lowest values recorded at Chatanika. The turbulence measurements coincided with the overturning of a 2.5-hr monochromatic gravity wave in a depth of 3 km at 85 km. The energy dissipation rates were estimated to be 3 mW/kg for the ensemble of waves and 18 mW/kg for the monochromatic wave. The MTeX observations reveal low levels of turbulence associated with low levels of gravity wave activity. In the light of other Arctic observations and model studies, these observations suggest that there may be reduced turbulence during disturbed winters.

An uncooled microbolometer-array thermal infrared camera has been incorporated into a remote sensing system for radiometric sky imaging. The radiometric calibration is validated and improved through direct comparison with spectrally integrated data from the Atmospheric Emitted Radiance Interferometer (AERI). With the improved calibration, the Infrared Cloud Imager (ICI) system routinely obtains sky images with radiometric uncertainty less than 0.5 W/(m(2 )sr) for extended deployments in challenging field environments. We demonstrate the infrared cloud imaging technique with still and time-lapse imagery of clear and cloudy skies, including stratus, cirrus, and wave clouds.