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RNA binding specificity of a Drosophila snRNP protein that shares sequence homology with mammalian U1-A and U2-B" proteins.
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We have characterized a recombinant Drosophila melanogaster RNA binding protein, D25, by virtue of its antigenic relationship to mammalian U1 and U2 small nuclear ribonucleoprotein (U snRNP) proteins. Sequence analysis revealed that D25 bears strong similarity to both the human U1 snRNP-A (U1-A) and U2 snRNP-B" (U2-B") proteins. However, at residues known to be critical for the RNA binding specificities of U1-A and U2-B" D25 sequence is more similar to U2-B". Using direct RNA binding assays D25 selected U1 RNA from either HeLa or Drosophila Kc cell total RNA. Furthermore, D25 bound U1 RNA when transfected into mammalian cells. Thus, D25 appears to be a Drosophila homolog of the mammalian U1-A protein, despite its sequence similarity to U2-B". 相似文献
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Large complex globular domains of type VII procollagen contribute to the structure of anchoring fibrils 总被引:18,自引:0,他引:18
G P Lunstrum L Y Sakai D R Keene N P Morris R E Burgeson 《The Journal of biological chemistry》1986,261(19):9042-9048
Type VII collagen, in the form of an antiparallel dimer, is a major protein component of anchoring fibrils. The ultrastructural appearance of these fibrils suggests that they may serve to anchor the basement membrane zone to the underlying connective tissue matrix. We report here the identification and initial characterization of Type VII procollagen, recovered from the media of epidermoid carcinoma cell cultures. Immunoblotting using monospecific antibodies to Type VII procollagen identifies a single, homogeneous band of at least Mr 320,000 following disulfide bond reduction. This chain contains 170 kDa of collagen triple helix and 150 kDa of non-helical domain at the carboxyl terminus. Pepsin digestion of this material yields Type VII collagen identical to that isolated from whole tissue and a series of quasi-stable peptides derived from the carboxyl-terminal region. Cell extracts contain procollagen chains identical in size to those secreted into the media. There is no evidence for processing of this material in cell culture. Partial purification by velocity sedimentation and transmission electron microscopic observation following rotary shadowing reveals both monomers (426 nm) and dimers (785 nm). Dimers are antiparallel and interact through 60-nm overlap, with amino-terminal globular domains present at the ends of the overlap. The multi-domain carboxyl-terminal region appears as three similar arms originating from a centralized globular region adjacent to the collagen helix. The carboxyl globular domain is present in whole tissue and may participate in the unique fibril form of this collagen. The amino-terminal globule may function in the antiparallel assembly of dimers. 相似文献
7.
Characterization of de novo duplications in eight patients by using fluorescence in situ hybridization with chromosome-specific DNA libraries.
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J Leana-Cox S Levin R Surana E Wulfsberg C L Keene L J Raffel B Sullivan S Schwartz 《American journal of human genetics》1993,52(6):1067-1073
Fluorescence in situ hybridization (FISH) with chromosome-specific DNA libraries was performed on samples from eight patients with de novo chromosomal duplications. In five cases, the clinical phenotype and/or cytogenetic evaluations suggested a likely origin of the duplicated material. In the remaining three cases, careful examination of the GTG-banding pattern indicated multiple possible origins; hybridization with more than one chromosome-specific library was performed on two of these cases. In all cases, FISH conclusively identified the chromosomal origin of the duplicated material. In addition, the hybridization pattern was useful in quantitatively delineating the duplication in two cases. 相似文献
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Two type XII-like collagens localize to the surface of banded collagen fibrils 总被引:8,自引:6,他引:2
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D R Keene G P Lunstrum N P Morris D W Stoddard R E Burgeson 《The Journal of cell biology》1991,113(4):971-978
Two recently identified collagen molecules, termed twelve-like A and twelve-like B (TL-A and TL-B) have properties similar to type XII collagen. These molecules have been localized in human and calf tissues by immunoelectron microscopy. The observations strongly suggest that both molecules are located along the surface of banded collagen fibers. The epitopes recognized by the antibodies are contained in large, nontriple-helical domains at one end of the collagen helix. The epitopes are visualized at a distance from the surface of the banded fibers roughly equal to the length of the nonhelical domains, suggesting that the nonhelical domains extend from the fibril, while the triple-helical domains are likely to bind directly to the fibril surface. Occasionally, both TL-A and TL-B demonstrate periodic distribution along the fibril surface. The period corresponds to the primary interband distance of the banded fibrils. Not all fibrils in a fiber bundle are labeled, nor is the labeling continuous along the length of labeled fibrils. Simultaneous labeling of TL-A and type VI collagen only rarely shows colocalization, suggesting that TL-A and TL-B do not mediate interactions between the type VI collagen beaded filaments and banded collagen fibrils. Also, interfibrillar distances are approximately equivalent in the presence and absence of these type XII-like molecules. While the results do not directly indicate a specific function for these molecules, the localization at the fibril surface suggests that they mediate interactions between the fibrils and other matrix macromolecules or with cells. 相似文献
9.
Kalinin: an epithelium-specific basement membrane adhesion molecule that is a component of anchoring filaments 总被引:56,自引:14,他引:42
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Basal keratinocytes attach to the underlying dermal stroma through an ultrastructurally unique and complex basement membrane zone. Electron-dense plaques along the basal surface plasma membrane, termed hemidesmosomes, appear to attach directly to the lamina densa of the basement membrane through fine strands, called anchoring filaments. The lamina densa is secured to the stroma through a complex of type VII collagen containing anchoring fibrils and anchoring plaques. We have identified what we believe is a novel antigen unique to this tissue region. The mAbs to this antigen localize to the anchoring filaments, just below the basal-dense plate of the hemidesmosomes. In cell culture, the antigen is deposited upon the culture substate by growing and migrating human keratinocytes. Addition of mAb to the cultures causes the cells to round and detach, but does not impair them metabolically. Skin fragments incubated with antibody extensively de-epithelialize. These findings strongly suggest that this antigen is intimately involved in attachment of keratinocytes to the basement membrane. This antigen was isolated from keratinocyte cultures by immunoaffinity chromatography. Two molecules are observed. The most intact species contains three nonidentical chains, 165, 155, and 140 kD linked by interchain disulfide bonds. The second and more abundant species contains the 165- and 140-kD chains, but the 155-kD chain has been proteolytically cleaved to 105 kD. Likewise, two rotary-shadowed images are observed. The larger of the two, presumably corresponding to the most intact form, appears as an asymmetric 107-nm-long rod, with a single globule at one end and two smaller globules at the other. The more abundant species, presumably the proteolytically cleaved form, lacks the distal small globule. We propose the name "kalinin" for this new molecule. 相似文献
10.
Summary During late spring, 1987, observations were made of nitrate and ammonium uptake in two regions of the Greenland Sea, the Arctic Front and the Polar Front. In the area of the Arctic Front, mixed layers were relatively deep (generally below 100m), and the 1% isolume averaged 35 m. Ambient nitrate concentrations were always greater than 6 M, whereas ammonium levels were always less than 0.6 M. Surface nitrate and ammonium specific uptake rates averages 4.4 and 2.3×10–3 h–1, respectively. The Polar Front generally coincided spatially with the location of the ice edge, and vertical mixed layers were shallow (pycnocline depth ranged from 8–14 m), and the 1 % isolume averaged 37 m. Nitrate concentrations were somewhat lower than in the Arctic Front, but remained above 3 M at all times. Ammonium levels reached 1.2 M. Nitrate and ammonium specific uptake rates at the surface averaged 4.8×10–3 and 10×10–3 h–1, respectively. Integrated water column f-ratios for the Arctic and Polar Front regions averaged 0.63 and 0.31, and the ammonium relative preference indices at all depths within each study area were always greater than 8, indicating that ammonium remained the preferred nitrogen source for phytoplankton. New production in the two regions was approximately equal, but the Polar Front had a substantially greater amount of regenerated production, and hence total production as well. Irradiance (and not nutrient concentration) seems to be the most important environmental factor in controlling nitrogen uptake. The spatial variability observed within the Greenland Sea suggest that inclusion of this region in global carbon models will require increased spatial resolution of both the models and the data included. 相似文献