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81.
Cytokines such as interferon-gamma (IFN-gamma), which utilize the well studied JAK/STAT pathway for nuclear signal transduction, are themselves translocated to the nucleus. The exact mechanism for the nuclear import of IFN-gamma or the functional role of the nuclear translocation of ligand in signal transduction is unknown. We show in this study that nuclear localization of IFN-gamma is driven by a simple polybasic nuclear localization sequence (NLS) in its COOH terminus, as verified by its ability to specify nuclear import of a heterologous protein allophycocyanin (APC) in standard import assays in digitonin-permeabilized cells. Similar to other nuclear import signals, we show that a peptide representing amino acids 95-132 of IFN-gamma (IFN-gamma(95-132)) containing the polybasic sequence 126RKRKRSR132 was capable of specifying nuclear uptake of the autofluorescent protein, APC, in an energy-dependent fashion that required both ATP and GTP. Nuclear import was abolished when the above polybasic sequence was deleted. Moreover, deletions immediately NH2-terminal of this sequence did not affect the nuclear import. Thus, the sequence 126RKRKRSR132 is necessary and sufficient for nuclear localization. Furthermore, nuclear import was strongly blocked by competition with the cognate peptide IFN-gamma(95-132) but not the peptide IFN-gamma(95-125), which is deleted in the polybasic sequence, further confirming that the NLS properties were contained in this sequence. A peptide containing the prototypical polybasic NLS sequence of the SV40 large T-antigen was also able to inhibit the nuclear import mediated by IFN-gamma(95-132). This observation suggests that the NLS in IFN-gamma may function through the components of the Ran/importin pathway utilized by the SV40 T-NLS. Finally, we show that intact IFN-gamma, when coupled to APC, was also able to mediate its nuclear import. Again, nuclear import was blocked by the peptide IFN-gamma(95-132) and the SV40 T-NLS peptide, suggesting that intact IFN-gamma was also transported into the nucleus through the Ran/importin pathway. Previous studies have suggested a direct intracellular role for IFN-gamma in the induction of its biological activities. Based on our data in this study, we suggest that a key intracellular site of interaction of IFN-gamma is the one with the nuclear transport mechanism that occurs via the NLS in the COOH terminus of IFN-gamma. 相似文献
82.
83.
KL Hsu CH Tsai FT Chiang HM Lo CD Tseng SG Ho YZ Tseng 《Canadian Metallurgical Quarterly》1997,80(12):1635-1637
Percutaneous balloon pericardiotomy is effective and less invasive for the treatment of recurrent pericardial effusion. This study suggests that the double-balloon method with 1 longer and 1 shorter balloon is the procedure of choice for percutaneous balloon pericardiotomy. 相似文献
84.
85.
What are the future directions of the field of nucleic acid crystallography? Although there have been many duplex structures determined, the sample is still relatively small. This is especially true if one wants to derive enough information about the relationships between sequence and structure. Indeed, there are data for all the possible 10 dimer steps, but for some steps it is very limited. If the structural code resides in trimers or tetrad steps then there is simply not enough data to do meaningful statistical analyses. So the first direction that needs to be explored is the determination of more structures with more varied sequences. The other noticeable thing about the data is the shortness of the strands. While it is probably true that attempts to crystallize very long sequences will not meet with success, the idea of crystallizing sequences engineered to fit together via sticky ends such as has been done for the CAP-DNA complex (Schultz et al., 1990) should give data about the behavior of much longer stretches of DNA. The question of the effects of environment on the structure of DNA continues to be a very important one to address since DNA is rarely alone. The preliminary data we have analysed from the current sample shows that the conformation of some steps are very sensitive to packing type. Numerous studies of the hydration around DNA shows that there is a real synergy between the hydration structure and the base conformation. More data will allow further quantitation of these observations. RNA structure is the next very exciting frontier. The emerging structures of duplexes with internal loops, the two hammerhead ribozyme structures and the group I intron ribozyme have given us a glimpse of the complexity and elegance of this class of molecules. With the technology now in place to allow the determination of the structures of these molecules, the expectation is that now we will see a large increase in the number of these structures in the NDB. 相似文献
86.
A Haaf L LeClaire G Roberts HM Kent TM Roberts M Stewart D Neuhaus 《Canadian Metallurgical Quarterly》1998,284(5):1611-1624
The major sperm protein (MSP) of Ascaris suum mediates amoeboid motility by forming an extensive intermeshed system of cytoskeletal filaments analogous to that formed by actin in many other amoeboid cells. MSP is a dimeric molecule that polymerizes to form non-polar filaments constructed from two helical subfilaments that wind round one another. Moreover, MSP filaments can interact with one another to form higher-order assemblies without requiring the range of accessory proteins usually employed in actin-based systems. A knowledge of how MSP polymerizes and forms the hierarchical series of helical MSP macromolecular assemblies is fundamental to understanding locomotion in these cells. Here we describe the solution structure of MSP dimers determined by NMR spectroscopy under conditions where MSP does not polymerize to form filaments. The solution structure is indistinguishable from that observed in putative MSP subfilament helices by X-ray crystallography, indicating that MSP polymerization is not accompanied by a major conformational change. We also show that the rate of MSP polymerization associated with movement of vesicles in an in vitro motility assay is enhanced by the presence of magnesium and manganese ions and use NMR to show that the primary residues that bind these ions are 24-25 and 83-86. These residues are distant from the interface formed between MSP dimers in subfilament helices, and so are probably not involved in this level of polymerization. Instead the manganese and magnesium ion binding appears to be associated with the assembly of subfilaments into filaments and their subsequent aggregation into bundles. 相似文献
87.
An immunoglobulin G of mouse was purified from sera by affinity chromatography in protein A. The rabbits whose sera were able to recognize the antigen injected by double immunodiffusion were immunized with this preparation. The antibodies were precipitated from the rabbit's serum and purified by ion exchange chromatography. This preparation was conjugated to fluorescin isothiocyanate according to the conventional technique. The conjugated obtained was evaluated with the reference strains of Parainfluenza virus 1, 2, 3; Adenovirus; respiratory syncytial virus; and influenza virus A and B, by an indirect immunofluorescence technique and HIV positive samples by flow citometry. Specific monoclonal antibodies were used in both cases. Clinical specimens of patients with acute respiratory infection were evaluated. 相似文献
88.
Stahl's ear is a congenital malformation of the auricle, which is uncommon in non-Oriental societies. A number of different treatment modalities have been suggested, many of which yield unpredictable results. We review current treatment modalities from the English literature and describe a novel, simple surgical method of repair. First, a helical rim incision is made to expose the abnormal third crus and upper pole of the ear. The third crus is excised as a narrow wedge of cartilage and posterior skin, and the defect is closed primarily. The free third crus cartilage is then grafted onto the scaphal cartilage in an anatomically correct position to form a superior crus. The anterior skin flap is redraped and bolstered over this graft. Only a very small scar on the helical rim remains visible anteriorly. 相似文献
89.
HM Cox 《Canadian Metallurgical Quarterly》1997,3(4):218-223
This paper describes one aspect of a study into the experiences in long-term healing of a community following the 1983 Ash Wednesday bushfire. Forty participants were interviewed, of whom 26 were residents and 14 disaster relief workers. The paper concentrates on the experiences of the latter, describing how they came to understand the bushfire affected the community and how they managed disaster work. For novices it was a profoundly difficult experience, for which they received little help and had to manage with whatever skill they drew on in their 'normal' working lives, mixed with a good deal of intuition. The paper suggests that health workers in vulnerable areas require preparation for a likely disaster; that 'outsiders' need to deal through existing community groups and individuals to gain access to those in need of their skills, and that they also require preparation for helping 'insiders' who are themselves victims of the catastrophe. 相似文献
90.
Reed Catherine L.; Lederman Susan J.; Klatzky Roberta L. 《Canadian Metallurgical Quarterly》1990,44(4):522
Three experiments involving a total of 75 college students investigated the role of size information in haptic classification of custom-made planar objects when size covaries with hardness, texture, or planar contour. Exp 1 showed facilitation of classification by redundant (RD) size and shape cues, indicating the coprocessing of size and shape. Exps 2 and 3 used a withdrawal paradigm: Classification trials began with 2 RD properties, and 1 was then held constant (withdrawn). Exp 2 showed that when size and shape were RD, withdrawal of either impaired responses, whereas when size was RD with texture or hardness, only size withdrawal had an effect. Exp 3 demonstrated that this size weighting was not restricted to a single procedure for exploration. (French abstract) (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献