bluuebunny/crossref_metadata_2025_split huggingface dataset:
GPR56/ADGRG1 activation induces IL‐6 production in melanoma cell via the Gα12/13‐ RhoA‐ROCK pathway
Authors: Huang Kuan‐Yeh , Chiang Nien‐Yi , Lin Hsi‐Hsien 
Published: June 2021

GPR56/ADGRG1, a member of the adhesion G protein‐coupled receptor family, is important for tumor progression. The role of GPR56 in melanoma development is unresolved. Here, we showed that GPR56 receptor ligation by immobilized CG4 mAb in melanoma cell enhanced the production of IL‐6, which promoted melanoma cell migration and invasion. siRNA‐mediated Gα12/13 knockdown significantly reduced CG4‐induced IL‐6 production in melanoma cell. Likewise, the IL‐6 level was decreased in melanoma cells expressing the dominant negative RhoA N19, treated with exoenzyme C3, or the ROCK inhibitors (Y27632 and H1152). We conclude that GPR56 activation‐induced IL‐6 production is mediated through the Gα12/13‐RhoA‐ROCK signaling pathway. IL‐6 in turn promotes melanoma cell migration and invasion. Our results indicate a critical role of GPR56 in melanoma progression.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural Basis of Glucocorticoid Receptor Recognition of Both Unmodified and Methylated Ancient Binding Sites, Precursors for a Modern Recognition Element
Authors: Liu Xu , Weikum Emily , Ortlund Eric 
Published: June 2021

DNA methylation plays crucial roles in gene silencing and development in eukaryotic cells. The most common DNA methylation involves the addition of a methyl group to a cytosine base in the context of a cytosine‐phosphate‐guanine (CpG) dinucleotide. Ancient genomes were enriched in CpG sites that may have served as transcription factor (TF) binding sites. These sites, through the process of methylation, deamination and subsequent mutation to TpG sites, may have maintained equal or enhanced TF binding. Recently, this has been described as a “mutator” system to produce genetic diversity during evolution. Here we test feasibility of this hypothesis by using an ancestral glucocorticoid receptor (GR) binding site (aGBS) that contains a CpG dinucleotide. We examine the binding of GR to ancestral unmodified, methylated and extant GBS and find two‐fold affinity increase upon the methylation and deamination. Additionally, this binding specificity is maintained from ancient steroid receptor AncSR2 to extant GR during the evolution. We also present the first structural studies of GR binding to unmodified and methylated aGBS. A methyl‐specific van der Walls contact with a Arg residue in the GR recognition helix is the molecular determinant of the binding specificity. Our study provides the first biochemical and structural evidence of high affinity binding for ancestral GR binding site containing methylated CpG, which could evolve to the extant, TpG‐containing site. This also indicates GR recognition of methylated CpG‐containing site is ancestral and offers new insight into evolution of steroid receptors and their binding sites.Support or Funding InformationThese studies were supported by National Institutes of Health [R01DK095750] and W.M. Keck Foundation Medical Research Grant to E. O. X.L was supported by an American Heart Association postdoctoral fellowship [17POST33660110].This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycine betaine accumulation in Avicennia germinans propagules and seedlings as a function of abiotic stress
Authors: Warren Jasmine N. , Day Zachary , Shor Audrey C. 
Published: June 2021

The black mangrove (Avicennia germinans) is native to the subtropical coasts of Florida and can tolerate moderate to high salinity environments. Propagules, germinated seeds that remain attached to the maternal plants until mature, are dispersed in shallow waters and transported through currents until washing ashore, and establishing roots on an appropriate substrate to initiate growth. Recently, the poleward expansion of A. germinans has been observed. The literature and findings in our laboratory suggest that A. germinans gains its salt and temperature tolerance in part by the modified amino acid, glycine betaine (GB). We sought to investigate the expression of GB in A. germinans as a function of abiotic stressors: high saline and low temperatures. The role of endogenous GB accumulation as a function of substrate condition and temperature was investigated in propagules. The accumulation of exogenous GB was measured in seedlings exposed to various saline and temperature environments. Propagules exposed to colder temperatures accumulated more GB (100–200 ug/umol) than moderate temperatures (50 ug/umol). Propagules exposed to 18 ppt saline accumulated more GB than those exposed to 36 ppt at 0°C. Propagules exposed to saline environments accumulated more GB at 0°C compared to sand and air samples. Significantly more propagules treated at 36 ppt developed into seedlings and demonstrated faster growth rates. Additionally, an exogenous application of 10 μM GB improved seedling survival after exposure to −2°C and high saline stress (60 ppt and 100 ppt). Preliminary findings demonstrate that APX and SOD enzymes are both present in the leaf samples after these stressors. These results further characterize the role GB contributes to the ability of A. germinans to acclimate following environmental stressors. Propagules express endogenous GB in response to abiotic stress, while exogenous application of GB may confer additional protection from continued stress. Subsequent investigations will further elucidate the roles APX and SOD may influence in the GB mechanism. The significance of these findings further characterize the mechanisms by which A. germinans survive high saline environments and colder temperatures.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein tyrosine phosphatase, receptor type B (PTPRB) inhibits brown adipocyte differentiation through regulation of VEGFR2 phosphorylation
Authors: Kim Won‐Kon , Han Baek‐Soo 
Published: June 2021

Brown adipocytes have an important role in the regulation of energy balance through uncoupling protein‐1 (UCP‐1)‐mediated nonshivering thermogenesis. Although brown adipocytes have been highlighted as a new therapeutic target for the treatment of metabolic diseases, such as obesity and type 2 diabetes in adult humans, the molecular mechanism underlying brown adipogenesis is not fully understood. We recently found that protein tyrosine phosphatase receptor type B (PTPRB) expression dramatically decreased during brown adipogenic differentiation. In this study, we investigated the functional roles of PTPRB and its regulatory mechanism during brown adipocyte differentiation. Ectopic expression of PTPRB led to a reduced brown adipocyte differentiation by suppressing the tyrosine phosphorylation of VEGFR2, whereas a catalytic inactive PTPRB mutant showed no effects on differentiation and phosphorylation. Consistently, the expression of brown adipocyte‐related genes, such as UCP‐1, PGC‐1α, PRDM16, PPAR‐γ and CIDEA, were significantly inhibited by PTPRB overexpression. Overall, these results suggest that PTPRB functions as a negative regulator of brown adipocyte differentiation through its phosphatase activity‐dependent mechanism and may be used as a target protein for the regulation of obesity and diabetes.Support or Funding InformationThis work was supported by grants from the KRIBB and from the Research Program (grants 2016R1C1B2013430, 2015M3A9B5030308, 2015M3A7B6027948, and 2015M3A9D7029882) through the National Research Foundation of Korea.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acid Phosphatase in Soil
Authors: Ayers Katherine Elizabeth , Fogle Emily J. 
Published: June 2021

Plants require phosphorus for growth, which they acquire from soil. This process is facilitated by the presence of enzymes, including acid phosphatases (AP). This enzyme catalyzes the cleavage of phosphate bonds, releasing the phosphate through hydrolysis. The degree of adsorption and activity of AP when adsorbed to soil is analyzed in order to determine how this environment effects the enzyme's action. A Bradford assay was used to determine concentration of AP and an activity assay utilizing fluorescent MUB to create a baseline characterization of AP in the presence of a clay, montmorillonite. Initial studies show that AP adsorbs to the clay and retains 34% activity. Future studies will look at the effect of time, concentration and temperature on activity levels.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Tumor Endothelial Marker 1 (TEM1/endosialin/CD248) Enhances Wound Healing Process by Cooperation with Platelet‐Derived Growth Factor (PDGF) Receptor
Authors: Hong Yi‐Kai 
Published: June 2021

Tumor endothelial marker 1 (TEM1), also named endosialin and CD248, is a type I transmembrane glycoprotein containing a C‐type lectin‐like domain. TEM1 is highly expressed on tumor‐associated fibroblasts, stromal cells, pericytes, and dermal fibroblasts. Dermal fibroblasts play a pivotal role in cutaneous wound healing which occurs in three dynamic and overlapping but distinct stages, including inflammatory phase, proliferation phase, and remodeling phase. However, the physiological function of TEM1 in wound healing is still unknown. In wound healing process both TEM1 and platelet‐derived growth factor receptor α (PDGFRα) expressions were highly upregulated in myofibroblasts. The cell activation, proliferation, and collagen deposition in granulation tissues were decreased and slower wound healing was found in TEM1‐deleted mice. In addition, the cell migration, adhesion and proliferation in NIH3T3 cells were decreased when TEM1 expression was knockdown by shRNA. The downstream signaling, mitogenic, and chemoattractive effects of platelet‐derived growth factor‐BB‐treated NIH3T3 cells were inhibited by knockdown of TEM1. TEM1 and PDGFRα were co‐localized in sub‐cellular organelles in fibroblasts and could be co‐immunoprecepitated. Based on these results we proposed that TEM1 in combination with PDGFRα plays a critical role in wound healing by enhancing the mitogenic and chemoattractive effects of platelet‐derived growth factor‐BB and collagen deposition in myofibroblasts.Support or Funding InformationThis work was supported by a grant from the Ministry of Science and Technology, Executive Yuan, Taiwan. (MOST 105‐2320‐B‐006 ‐039 ‐MY3 to Hua‐Lin Wu)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiac Nav1.5 Channel is Regulated by LITAF
Authors: Turan Nilufer Nermin , Moshal Karni S. , Roder Karim , Xie An , Lu Yichun , Werdich Andreas , MacRae Calum , Koren Gideon 
Published: June 2021

BackgroundVoltage‐gated Na+ channels (Nav1.5) are highly expressed in the heart, trigger electrical impulses and contribute to the action potential (AP). Genome‐wide association studies suggest that lipopolysaccharide‐induced tumor necrosis factor‐alpha factor (LITAF) is a likely modifier of the QT interval. LITAF is an adapter molecule for members of the Nedd4‐like family of E3 ubiquitin ligases and is involved in endosomal trafficking. Nedd4‐2 is responsible for ubiquitination of Nav1.5 resulting in its degradation.GoalWe postulate that LITAF inhibits the ubiquitination activity of Nedd4‐2 or restricts the access of Nedd4‐2 to the Nav1.5 complex on the plasma membrane, thus inhibiting channel ubiquitination and degradation via the lysosome pathway. Hence, a loss‐of‐function of LITAF (as seen in heart failure) will lead to increased degradation of voltage‐gated sodium channels and impair conduction in the heart.MethodsCultured 3‐week old rabbit cardiomyocytes (3wRbCM) were infected with adenovirus (MOI 10) expressing HA‐tagged LITAF or GFP. HEK cells stably expressing Nav1.5 were transfected with expression plasmids for HA‐LITAF, its PXY double mutant (HA‐LITAF‐Y23A‐Y61A) and/or Nedd4‐2. Patch clamping was performed to measure INa. Western blotting and co‐immunoprecipitation were used to measure protein levels and detect protein‐protein interactions, respectively.ResultsIn 3wRbCM, Nav1.5 channel expression and Nav1.5 current were significantly increased by LITAF. These effects could be recapitulated in HEK cells stably expressing Nav1.5. However, the PXY mutations in LITAF abolished the LITAF‐dependent increase in Nav1.5 protein and encoded current. Immunoprecipitations demonstrated a physical interaction between LITAF and Nav1.5 or NEDD4‐2. Additionally, LITAF suppressed the expression of both endogenous and co‐expressed NEDD4‐2.ConclusionsLITAF is a novel regulator of cardiac Nav1.5 and action potential duration, likely by affecting Nedd4‐2 levels via increasing its autoubiquitination activity in the cell.Support or Funding Information

International Postdoctoral Research Scholarship Programme (2219) by The Scientific and Technological Research Council of Turkey.
The National Institutes of Health [R01‐HL‐096669, T32HL094300, R01HL110791, R01HL134706].
The Rhode Island Foundation [701‐7131778].
This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early growth response protein‐1 (Egr‐1) expression by hydrogen peroxide (H2O2) is mediated via c‐Src and PKB‐dependent CREB signaling events in vascular smooth muscle cells (VSMC)
Authors: Truong Vanessa , Rondeau Vincent , Jain Ashish , Srivastava Ashok K. 
Published: June 2021

Increased generation of reactive oxygen species (ROS) is believed to play a key role in the pathophysiology of cardiovascular diseases. Excessive growth and proliferation of vascular smooth muscle cells (VSMC) have been suggested to be major contributors to vascular dysfunction. A potential involvement of early growth response protein‐1 (Egr‐1), a zinc‐finger transcription factor, in the development of vascular injury has been suggested. Recent studies have shown that H2O2, one of the most stable ROS, increases Egr‐1 expression in VSMC, however, signaling events leading to H2O2‐induced Egr‐1 expression are not fully understood. Here, we report that H2O2 increased Egr‐1 expression in a time and dose‐dependent fashion in A10 VSMC and pharmacological blockade of insulin‐like growth factor‐1 receptor (IGF‐1R) and c‐Src, by AG1024 and PP2 respectively, abolished H2O2‐induced Egr‐1 expression while AG1478, an inhibitor of the epidermal growth factor receptor (EGFR), and PP3, the inactive analog of PP2, had no effect. H2O2 also stimulated the phosphorylation of PKB, and siRNA‐induced silencing of c‐Src resulted in a significant reduction in PKB phosphorylation as well Egr‐1 expression. In addition, phosphorylation of cyclic AMP response element binding protein (CREB) was also enhanced by H2O2, however, inhibition of PI3K/PKB by wortmannin and SC‐66 attenuated this phosphorylation, as well as suppressed protein and mRNA levels of Egr‐1 induced by H2O. In summary, our data suggests that H2O2 provokes Egr‐1 expression through the activation of upstream tyrosine kinases IGF‐1R and c‐Src as well as PKB and CREB‐dependent signaling events in VSMC.Support or Funding Information(Supported by CIHR)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Revealing Subcellular Redox Dynamics with Multiplex Imaging of Compartment‐Specific Redox Probes
Authors: Tantama Mathew , Norley Jacob , Radhakrishnan Saranya , Rajendran Megha 
Published: June 2021

Reactive oxygen species (ROS), redox enzymes, and redox buffers are central to the propagation of redox signals and oxidative stress between cells in the tissue environment and between organelles in the cellular environment. For example, hydrogen peroxide generated in mitochondria can alter the thiol oxidation states on proteins to alter their function, and subsequently redox active biomolecules such as glutathione can act as reducing equivalents in the face of oxidative stress. In some cases, it has become apparent that ROS mediate physiologically important signaling across compartments, such as in mitonuclear retrograde signaling. However, it has remained technically challenging to quantitatively correlate compartment‐specific redox dynamics with subcellular resolution within live single cells. In order to address this challenge, we are developing genetically‐encoded biosensors that enable the visualization of redox dynamics simultaneously within multiple compartments within the same cell. Our approach utilizes Förster‐type resonance energy transfer in a spectral relay strategy to extend the fluorescence emission of the redox‐sensitive green fluorescent protein (roGFP) into orange and red emission wavelengths. Furthermore, we are developing multiplexed, multiphoton imaging methods to study redox coupling between organelles and compartment‐specific dynamics in thick neural tissues.Support or Funding InformationWe gratefully acknowledge support from the Ralph W. and Grace M. Showalter Trust, the Purdue Research Foundation, and from National Institutes of Health grants R21 NS092010 and R21 EY026425.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Converting Nature's Switches Into Scientists' Tools: How Biophysical Insights Lay The Foundation For Artificial Control Of Protein Activity
Authors: Gardner Kevin H. , Dikiy Igor , Edupuganti Uthama , Orth Elizabeth , Abzalimov Rinat 
Published: June 2021

Environmental cues regulate many biological processes, coordinating cellular pathways to respond to changing conditions. Such regulation is often initiated by sensory protein domains which expand their chemical repertoire by using small molecule ligands to convert environmentally‐triggered changes into altered protein/protein interactions. Several families of these domains have evolved with remarkable diversity in their inputs and outputs. Using a combination of biophysics, biochemistry and synthetic chemistry, we seek to gain insight into the mechanistic controls of such environmental sensing domains for both fundamental understanding and subsequent artificial control.Here I will discuss examples of our work exploring how the Light‐Oxygen‐Voltage (LOV) protein domains convert changes in a common blue light stimulus into control of a wide range of output functions. Such photosensory domains utilize flavin chromophores and a photochemically‐generated protein/flavin adduct as a trigger for the allosteric control of effectors as diverse as kinases to DNA‐binding domains. While high‐resolution structures of inhibited LOV proteins have provided insights into certain regulatory principles, it has been difficult to apply similar techniques to spontaneously‐activated “dark noise” and photoactivated conformations that are essential for a full understanding of control. We have tackled this challenge by combining data from several biophysical approaches, including HDX‐MS and EPR spectroscopy, letting us observe and characterize these states. This information has shown several commonalities of LOV signaling in diverse classes of sensory proteins, and further, develop structure‐based mutations to shift these proteins among these states. Doing so, we have taken advantage of this mechanistic understanding to develop the artificial regulation of such systems, both in vitro and in living cells. Taken together, our work provides an integrated view of a fascinating class of natural switches and suggests routes by which these can be manipulated to achieve desired therapeutic and/or technological outcomes.Support or Funding InformationSupport from NIH R01 GM106239 is gratefully acknowledged.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Growth of Fusarium oxysporum Spore Isolates from Potato is Inhibited by 1,4‐dimethylnaphthalene
Authors: Adams Rachel , Campbell Michael 
Published: June 2021

Prolongation of storage via temporary meristematic sprout inhibition has effectively been carried out in tubers after brief exposure to 1,4‐dimethylnaphthalene (DMN), a compound naturally abundant in the tissues of dormant potatoes. Not only does DMN suppress meristematic growth, but studies have shown that DMN greatly impacts the microbiome on the surface of potatoes, especially with regards to species diversity. Among the diverse microbiome population is Fusarium oxysporum, a particular strain of pathogenic fungus responsible for the plant disease fusarium wilt. F. oxysporum was successfully isolated from the surface of a tuber using the fusarium selective media Nash‐Snider, to which cultures were transferred and maintained on nutritionally complex potato dextrose agar plates. DNA sequencing using selective primers NL1, NL2, ITS1F, and ITS2R confirmed the fungus to be F. oxysporum. A single spore isolation technique was utilized to record the number and growth response of spores treated with DMN vs spores treated with water (control). Single spore isolate plates were contained in vacuum sealed chambers and incubated at a constant 22C while exposed to 7.5ul DMN via aerosol dispersion for four days. After a day of rest, the spores on each plate were measured and counted. The results concluded spores exposed to DMN showed slower diameter growth than those treated with water. Knowing the effect of DMN on F. oxysporum is critical in controlling and preventing harvest loss brought on by fusarium wilt. In an attempt to gain more insight on the specificities of the DMN‐fungal conundrum, further studies are still being conducted.Support or Funding InformationSupported in part by a grant from the 1,4‐Group, Meridian, IDThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.