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991.
Meisner NC Hackermüller J Uhl V Aszódi A Jaritz M Auer M 《Chembiochem : a European journal of chemical biology》2004,5(10):1432-1447
Approximately 3 000 genes are regulated in a time-, tissue-, and stimulus-dependent manner by degradation or stabilization of their mRNAs. The process is mediated by interaction of AU-rich elements (AREs) in the mRNA's 3'-untranslated regions with trans-acting factors. AU-rich element-controlled genes of fundamentally different functional relevance depend for their activation on one positive regulator, HuR. Here we present a methodology to exploit this central regulatory process for specific manipulation of AU-rich element-controlled gene expression at the mRNA level. With a combination of single-molecule spectroscopy, computational biology, and molecular and cellular biochemistry, we show that mRNA recognition by HuR is dependent on the presentation of the sequence motif NNUUNNUUU in single-stranded conformation. The presentation of the HuR binding site in the mRNA secondary structure appears to act analogously to a regulatory on/off switch that specifically controls HuR access to mRNAs in cis. Based on this finding we present a methodology for manipulating ARE mRNA levels by actuating this conformational switch specifically in a target mRNA. Computationally designed oligonucleotides (openers) enhance the NNUUNNUUU accessibility by rearranging the mRNA conformation. Thereby they increase in vitro and endogenous HuR-mRNA complex formation which leads to specific mRNA stabilization (as demonstrated for TNFalpha and IL-2, respectively). Induced HuR binding both inside and outside the AU-rich element promotes functional IL-2 mRNA stabilization. This opener-induced mRNA stabilization mimics the endogenous IL-2 response to CD28 stimulation in human primary T-cells. We therefore propose that controlled modulation of the AU-rich element conformation by mRNA openers or closers allows message stabilization or destabilization in cis to be specifically triggered. The described methodology might provide a means for studying distinct pathways in a complex cellular network at the node of mRNA stability control. It allows ARE gene expression to be potentially silenced or boosted. This will be of particular value for drug-target validation, allowing the diseased phenotype to ameliorate or deteriorate. Finally, the mRNA openers provide a rational starting point for target-specific mRNA stability assays to screen for low-molecular-weight compounds acting as inhibitors or activators of an mRNA structure rearrangement. 相似文献
992.
We tested the ability of male slow-worms, Anguis fragilis, a limbless anguid lizard with secretive, semifossorial habits, to detect chemical associated with conspecifics by using a T-maze in the laboratory. Male slow-worms discriminated conspecific male and female scent deposits. Males selected the arm with female scent, suggesting that scent deposits may be used to locate potential mates. Also, male slow-worms did not avoid the chemicals of other males, suggesting that they are not territorial. However, males discriminated their own scent from those of other males, and spent more time exploring the arm with other male scent, which suggests that scent marks may bear information that could be used in future intrasexual social contexts. We conclude that discrimination of conspecifics based on scents may be more widespread than previously expected among lizards inhabiting visually restricted environments. 相似文献
993.
A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents 总被引:4,自引:0,他引:4
Thermostable variants of the Class II fructose bisphosphate aldolase have been isolated following four rounds of directed evolution using DNA shuffling of the fda genes from Escherichia coli and Edwardsiella ictaluri. Variants from all four generations of evolution have been purified and characterized. The variants show increased thermostability with no loss of catalytic function at room temperature. The temperature at which 50% of the initial enzyme activity is lost after incubation for 10 min (T50) of the most stable variant, 4-43D6, is increased by 11-12 degrees C over the wild-type enzymes and the half-life of activity at 53 degrees C is increased approximately 190-fold. In addition, variant 4-43D6 shows increased stability to treatment with organic solvents. DNA sequencing of the evolved variants has identified the mutations which have been introduced and which lead to increased thermostability, and the role of the mutations introduced is discussed. 相似文献
994.
Dai J Park G Perry JL Il'ichev YV Bow DA Pritchard JB Faucet V Pfohl-Leszkowicz A Manderville RA Simon JD 《Accounts of chemical research》2004,37(11):874-881
Ochratoxins are a class of naturally occurring compounds produced by several fungi. The most toxic is ochratoxin A (OTA), and occurrence of some human nephropathies and tumors correlate with enhanced OTA exposure. In this Account, the following areas are examined: molecular details of the binding of OTA to human serum albumin (HSA), the influences of binding to HSA on the trans-port of OTA across epithelial cell membranes by organic anion transport proteins, the oxidative activation of OTA, and the formation of OTA adducts with biological molecules. These studies are beginning to provide a detailed chemical model for the trans-port, accumulation, and genotoxic and carcinogenic effects of OTA. 相似文献
995.
The bolas spider, Mastophora hutchinsoni, attracts adult males of four species of moths by aggressively mimicking their sex pheromone. Here, we report the identification of two sex pheromone components of one of these species, the bronzed cutworm, Nephelodes minians. Simultaneous gas chromatographic (flame ionization detection) and electroantennographic detection (EAD) analysis of extracts of the sex pheromone glands of female N. minians indicated two components eliciting strong EAD responses from a conspecific male antenna that corresponded in retention time with (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-11-hexadecenyl acetate (Z11-16:OAc), using both polar and nonpolar columns. Mass spectra of these two peaks were identical to those of synthetic Z11-16:Ald and Z11-16:OAc. The double-bond positions were confirmed by GC-MS analysis of dimethyl disulfide derivatives of the putative pheromone components. In a field test, a 5:1 blend of Z11-16:Ald to Z11-16:OAc attracted male N. minians. Overall, there is no overlap in the chemical constituents of the sex pheromones of three moth species preyed upon by this bolas spider for which pheromones have now been identified. These studies suggest that bolas spiders are versatile in their ability to synthesize semiochemicals for different prey species. This versatility may be advantageous to a predator that has an otherwise extremely specialized hunting tactic. 相似文献
996.
997.
998.
研究了全球肥皂厂家或研发实验室采用的评定方法,这些评定的目的是为了确保其满足生产标准,并最终保证消费者对他们购买到的肥皂产品感到满意。一般来说,评定方法包括分析化学检测(评定原材料和成品)、材料学法(评定产品结构)以及消费者可见属性的直接测试(例如泡沫、湿皂触感和磨损率)。这些检测可在实验室中进行或通过消费者评估进行。 相似文献
999.
π-Conjugated organic molecules represent an attractive platform for the design and fabrication of a wide range of nano- and microstructures for use in organic optoelectronics. The desirable optical and electrical properties of π-conjugated molecules for these applications depend on their primary molecular structure and their intermolecular interactions such as molecular packing or ordering in the condensed states. Because of the difficulty in satisfying these rigorous structural requirements for photoluminescence and charge transport, the development of novel high-performance π-conjugated systems for nano-optoelectronics has remained a challenge. This Account describes our recent discovery of a novel class of self-assembling π-conjugated organic molecules with a built-in molecular elastic twist. These molecules consist of a cyano-substituted stilbenic π-conjugated backbone and various terminal functional groups, and they offer excellent optical, electrical, and self-assembly properties for use in various nano-optoelectronic devices. The characteristic "twist elasticity" behavior of these molecules occurs in response to molecular interactions. These large torsional or conformational changes in the cyanostilbene backbone play an important role in achieving favorable intermolecular interactions that lead to both high photoluminescence and good charge carrier mobility in self-assembled nanostructures. Conventional π-conjugated molecules in the solid state typically show concentration (aggregation) fluorescence quenching. Initially, we describe the unique photoluminescence properties, aggregation-induced enhanced emission (AIEE), of these new cyanostilbene derivatives that elegantly circumvent these problems. These elastic twist π-conjugated backbones serve as versatile scaffolds for the preparation of well-defined patterned nanosized architectures through facile self-assembly processes. We discuss in particular detail the preparation of 1D nanowire structures through programmed self-assembly. This Account describes the importance of utilizing AIEE effects to explore optical device applications, such as organic semiconducting lasers (OSLs), optical memory, and sensors. We demonstrate the rich electronic properties, including the electrical conductivity, field-effect carrier mobility, and electroluminescence of highly crystalline 1D nanowire and coaxial donor-acceptor nanocable structures composed of elastic twist π-conjugated molecules. The electronic properties were measured using various techniques, including current-voltage (I-V), conducting-probe atomic force microscopy (CP-AFM), and space-charge-limited-current (SCLC) measurements. We prepared and characterized several electronic device structures, including organic field-effect transistors (OFETs) and organic light-emitting field-effect transistors (OLETs). 相似文献
1000.