Intermolecular disulfide bond influences unphosphorylated STAT3 dimerization and function

Signal transducer and activator of transcription 3 (STAT3) is a transcription factor activated by the phosphorylation of tyrosine 705 in response to many cytokines and growth factors. Recently, the roles for unphosphorylated STAT3 (U-STAT3) have been described in response to cytokine stimulation, in cancers, and in the maintenance of heterochromatin stability. It has been reported that U-STAT3 dimerizes, shuttles between the cytoplasm and nucleus, and binds to DNA, thereby driving genes transcription. Although many reports describe the active role of U-STAT3 in oncogenesis in addition to phosphorylated STAT3, the U-STAT3 functional pathway remains elusive.In this report, we describe the molecular mechanism of U-STAT3 dimerization, and we identify the presence of two intermolecular disulfide bridges between Cys367 and Cys542 and Cys418 and Cys426, respectively. Recently, we reported that the same cysteines contribute to the redox regulation of STAT3 signaling pathway both in vitro and in vivo The presence of these disulfides is here demonstrated to largely contribute to the structure and the stability of U-STAT3 dimer as the dimeric form rapidly dissociates upon reduction in the S-S bonds. In particular, the Cys367-Cys542 disulfide bridge is shown to be critical for U-STAT3 DNA-binding activity. Mutation of the two Cys residues completely abolishes the DNA-binding capability of U-STAT3. Spectroscopic investigations confirm that the noncovalent interactions are sufficient for proper folding and dimer formation, but that the interchain disulfide bonds are crucial to preserve the functional dimer. Finally, we propose a reaction scheme of U-STAT3 dimerization with a first common step followed by stabilization through the formation of interchain disulfide bond

Network and atomistic simulations unveil the structural determinants of mutations linked to retinal diseases

A number of incurable retinal diseases causing vision impairments derive from alterations in visual phototransduction. Unraveling the structural determinants of even monogenic retinal diseases would require network-centered approaches combined with atomistic simulations. The transducin G38D mutant associated with the Nougaret Congenital Night Blindness (NCNB) was thoroughly investigated by both mathematical modeling of visual phototransduction and atomistic simulations on the major targets of the mutational effect. Mathematical modeling, in line with electrophysiological recordings, indicates reduction of phosphodiesterase 6 (PDE) recognition and activation as the main determinants of the pathological phenotype. Sub-microsecond molecular dynamics (MD) simulations coupled with Functional Mode Analysis improve the resolution of information, showing that such impairment is likely due to disruption of the PDEgamma binding cavity in transducin. Protein Structure Network analyses additionally suggest that the observed slight reduction of theRGS9-catalyzed GTPase activity of transducin depends on perturbed communication between RGS9 and GTP binding site. These findings provide insights into the structural fundamentals of abnormal functioning of visual phototransduction caused by a missense mutation in one component of the signaling network. This combination of network-centered modeling with atomistic simulations represents a paradigm for future studies aimed at thoroughly deciphering the structural determinants of genetic retinal diseases. Analogous approaches are suitable to unveil the mechanism of information transfer in any signaling network either in physiological or pathological conditions

Hydrothermal depolymerization of biorefinery lignin-rich streams: Influence of reaction conditions and catalytic additives on the organic monomers yields in biocrude and aqueous phase

Hydrothermal depolymerization of lignin-rich streams (LRS) from lignocellulosic ethanol was successfully carried out in a lab-scale batch reactors unit. A partial depolymerization into oligomers and monomers was achieved using subcritical water as reaction medium. The influence of temperature (300–350–370 ◦C) and time (5–10 minutes) was investigated to identify the optimal condition on the monomers yields in the lighter biocrude (BC1) and aqueous phase (AP) fractions, focusing on specific phenolic classes as well as carboxylic acids and alcohols. The effect of base catalyzed reactions (2–4 wt. % of KOH) was compared to the control tests as well as to acid-catalyzed reactions obtained with a biphasic medium of supercritical carbon dioxide (sCO2) and subcritical water. KOH addition resulted in enhanced overall depolymerization without showing a strong influence on the phenolic generation, whereas sCO2 demonstrated higher phenolic selectivity even though no effect was observed on the overall products mass yields. In conclusion, a comparison between two different biocrude collection procedures was carried out in order to understand how the selected chemical extraction mode influences the distribution of compounds between BC1 and AP

Hydrothermal depolymerization of biorefinery lignin-rich streams: Influence of reaction conditions and catalytic additives on the organic monomers yields in biocrude and aqueous phase

Hydrothermal depolymerization of lignin-rich streams (LRS) from lignocellulosic ethanol was successfully carried out in a lab-scale batch reactors unit. A partial depolymerization into oligomers and monomers was achieved using subcritical water as reaction medium. The influence of temperature (300–350–370 °C) and time (5–10 minutes) was investigated to identify the optimal condition on the monomers yields in the lighter biocrude (BC1) and aqueous phase (AP) fractions, focusing on specific phenolic classes as well as carboxylic acids and alcohols. The effect of base catalyzed reactions (2–4 wt. % of KOH) was compared to the control tests as well as to acid-catalyzed reactions obtained with a biphasic medium of supercritical carbon dioxide (sCO2) and subcritical water. KOH addition resulted in enhanced overall depolymerization without showing a strong influence on the phenolic generation, whereas sCO2 demonstrated higher phenolic selectivity even though no effect was observed on the overall products mass yields. In conclusion, a comparison between two different biocrude collection procedures was carried out in order to understand how the selected chemical extraction mode influences the distribution of compounds between BC1 and AP

Predictors of children's secondhand smoke exposure at home: a systematic review and narrative synthesis of the evidence

BACKGROUND: Children's exposure to secondhand smoke (SHS) has been causally linked to a number of childhood morbidities and mortalities. Over 50% of UK children whose parents are smokers are regularly exposed to SHS at home. No previous review has identified the factors associated with children's SHS exposure in the home. AIM: To identify by systematic review, the factors which are associated with children's SHS exposure in the home, determined by parent or child reports and/or biochemically validated measures including cotinine, carbon monoxide or home air particulate matter. METHODS: Electronic searches of MEDLINE, EMBASE, PsychINFO, CINAHL and Web of Knowledge to July 2014, and hand searches of reference lists from publications included in the review were conducted. FINDINGS: Forty one studies were included in the review. Parental smoking, low socioeconomic status and being less educated were all frequently and consistently found to be independently associated with children's SHS exposure in the home. Children whose parents held more negative attitudes towards SHS were less likely to be exposed. Associations were strongest for parental cigarette smoking status; compared to children of non-smokers, those whose mothers or both parents smoked were between two and 13 times more likely to be exposed to SHS. CONCLUSION: Multiple factors are associated with child SHS exposure in the home; the best way to reduce child SHS exposure in the home is for smoking parents to quit. If parents are unable or unwilling to stop smoking, they should instigate smoke-free homes. Interventions targeted towards the socially disadvantaged parents aiming to change attitudes to smoking in the presence of children and providing practical support to help parents smoke outside the home may be beneficial