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The positive-acting global sulfur regulatory protein, CYS3, of Neurospora crassa turns on the expression of a family of unlinked structural genes that encode enzymes of sulfur catabolism. CYS3 contains a leucine zipper and an adjacent basic region (b-zip), which together constitute a bipartite sequence-specific DNA-binding domain. Specific anti-CYS3 antibodies detected a protein of the expected size in nuclear extracts of wild-type Neurospora under conditions in which the sulfur circuit is activated. The CYS3 protein was not observed in cys-3 mutants. Nuclear extracts of wild type, but not cys-3 mutants, also showed specific DNA-binding activity identical to that obtained with a CYS3 protein expressed in Escherichia coli. A truncated CYS3 protein that contains primarily the b-zip domain binds to DNA with high specificity and affinity in vitro, yet fails to activate gene expression in vivo, and instead inhibits the function of the wild-type CYS3 protein. Amino-terminal, carboxyterminal, and internal deletions as well as alanine scanning mutagenesis were employed to identify regions of the CYS3 protein that are required for its trans-activation function. Regions of CYS3 carboxy terminal to the b-zip motif are not completely essential for function although loss of an alanine-rich region results in decreased activity. All deletions amino terminal to the b-zip motif led to a complete loss of CYS3 function. Alanine scanning mutagenesis demonstrated that an unusual prolinerich domain of CYS3 appears to be very important for function and is presumed to constitute an activation domain. It is concluded that CYS3 displays nuclear localization and positive autogenous control in Neurospora and functions as a trans-acting DNA-binding protein. 相似文献
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SR DeMeester GA Patterson RS Sundaresan JD Cooper 《Canadian Metallurgical Quarterly》1998,115(3):681-688
Methanol-induced conformational transitions of hen egg white lysozyme were investigated with a combined use of far- and near-UV CD and NMR spectroscopies, ANS binding and small-angle X-ray scattering. Addition of methanol induced no global change in the native conformation itself, but induced a transition from the native state to the denatured state which was highly cooperative, as shown by the coincidence of transition curves monitored by the far- and near-UV CD spectroscopy, by isodichroic points in the far- and near-UV CD spectra and by the concomitant disappearance of individual 1H NMR signals of the native state. The ANS binding experiments could detect no intermediate conformer similar to the molten globule state in the process of the methanol denaturation. However, at high concentration of methanol, e.g., 60% (v/v) methanol/water, a highly helical state (H) was realized. The H state had a helical content much higher than the native state, monitored by far-UV CD spectroscopy, and had no specific tertiary structure, monitored both by near-UV CD and NMR spectroscopy. The radius of gyration in the H state, 24.9 angstroms, was significantly larger than that in the native state (15.7 angstroms). The Kratky plot for the H state did not show a clear peak and was quite similar to that for the urea-denatured state, indicating a complete lack of globularity. Thus we conclude that the H state has a considerably expanded, flexible broken rod-like conformation which is clearly distinguishable from the "molten globule" state. The stability of both N and H states depends on pH and methanol concentration. Thus a phase diagram involving N and H was constructed. 相似文献
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AD Lelianov IuG Novikov GA Rusanov NE Siniavskaia 《Canadian Metallurgical Quarterly》1976,116(4):24-28
In experiments on 50 dogs with toxic acute edema of the lung, induced with intravenous injection of 0.1% silver nitrate, the authors have studied the efficacy of accessory artificial circulation and "conservative" therapy. During the perfusion a discharge of the right portions, adequate extracorporeal gas metabolism, normalization of blood gas and acid-base balance were noted; an intensity of pulmonary edema is descreased. An intensive therapy for pulmonary edema was found to be more effective in association of "conservative" treatment with venoarterial perfusion and blood oxygenation. 相似文献
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主蒸汽压力是电厂热工过程控制中的重要参数,由于锅炉燃烧工况变化较大,主汽压对象模型具有不确定性,常规控制方法很难得到预期的控制效果。为提高系统控制品质,提出了主汽压系统的H∞混合灵敏度设计方法,给出了选择加权函数的具体方法,采用MATLAB软件进行仿真计算,得到H∞最优控制器。仿真结果表明,在对象模型参数发生较大变化时,与常规PID调节方法相比,H∞最优控制器能使主汽压控制系统具有良好的鲁棒稳定性和动态性能。 相似文献
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Marijuana, Cannabis sativa, elicits a variety of effects in experimental animals and humans. Delta-9-tetrahydrocannabinol (THC) is the major psychoactive component in marijuana. This substance has been shown, also, to be immunosuppressive and to decrease host resistance to bacterial, protozoan, and viral infections. Macrophages, T lymphocytes, and natural killer cells appear to be major targets of the immunosuppressive effects of THC. Definitive data which directly link marijuana use to increased susceptibility to infection in humans currently is unavailable. However, cumulative reports indicating that THC alters resistance to infection in vitro and in a variety of experimental animals support the hypothesis that a similar effect occurs in humans. 相似文献
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