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71.
Identification and analysis of a retinoblastoma binding motif in the replication protein of a plant DNA virus: requirement for efficient viral DNA replication. 总被引:22,自引:3,他引:19
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Geminiviruses are plant DNA viruses with small genomes whose replication, except for the viral replication protein (Rep), depends on host proteins and, in this respect, are analogous to animal DNA tumor viruses, e.g. SV40. The mechanism by which these animal viruses create a cellular environment permissive for viral DNA replication involves the binding of a virally encoded oncoprotein, through its LXCXE motif, to the retinoblastoma protein (Rb). We have identified such a LXCXE motif in the Rep protein of wheat dwarf geminivirus (WDV) and we show its functional importance during viral DNA replication. Using a yeast two-hybrid system we have demonstrated that WDV Rep forms stable complexes with p130Rbr2, a member of the Rb family of proteins, and single amino acid changes within the LXCXE motif abolish the ability of WDV Rep to bind to p130Rbr2. The LXCXE motif is conserved in other members of the same geminivirus subgroup. The presence of an intact Rb binding motif is required for efficient WDV DNA replication in cultured wheat cells, strongly suggesting that one of the functions of WDV Rep may be the linking between viral and cellular DNA replication cycles. Our results point to the existence of a Rb-like protein(s) in plant cells playing regulatory roles during the cell cycle. 相似文献
72.
Allergenic and antigenic proteins released in the apertural sporoderm during the activation process in grass pollen grains 总被引:2,自引:0,他引:2
Jesús Márquez Juan A. Seoane-Camba María Suárez-Cervera 《Sexual plant reproduction》1997,10(5):269-278
A combination of transmission electron microscopy with immunocytochemical methods was used to localize antigenic and allergenic proteins during the maturation and activation processes of Poaceae pollen grains. The intine undergoes a series of modifications that play a decisive role in these processes. Allergenic and antigenic proteins were detected particularly in the intine of activated in vitro grass pollen grains. Labelling of antigenic proteins was more abundant and less specific than that of allergenic proteins. At the time of hydration, the operculum was lifted up, the intine was swollen and labelling of allergenic proteins appeared highly localized in the Zwischenkörper. No significant labelling was observed when the Zwischenkörper gelatinized. Immunolocalization of allergenic proteins in the activated Zwischenkörper indicated the presence of proteins related to activation of the pollen grains. This confirms that the intine function is involved in the processes of pollen tube formation and fertilization, and also suggests the possible mechanism activated in the pollen grains when allergenic proteins reach the mucosa of sensitive subjects. 相似文献
73.
The power of heteronuclear NMR spectroscopy to study macromoleculesand their complexes has been amply demonstrated over the last decade. Theobstacle to routinely applying these techniques to the study of DNA has beenthe synthesis of 13C,15N-labeled DNA. Here wepresent a simple and efficient method to generate isotope-labeled DNA forNMR studies that is as easy as that for isotope labeling of RNA. The methodwas used to synthesize a uniformly13 C,15N-labeled 32-nucleotide DNA that binds tohuman basic fibroblast growth factor with high affinity and specificity.Isotope-edited experiments were applied to the13 C,15N-labeled DNA bound to unlabeled protein,and the 13 C,15N-labeled DNA was also examined incomplex with 15N-labeled protein. The NMR experiments showthat the DNA adopts a well-defined stable structure when bound to theprotein, and illustrate the potential of13 C,15N-labeled DNA for structural studies ofDNA–protein complexes. 相似文献
74.
75.
Molecular structure of the halogenated anti-cancer drug iododoxorubicin complexed with d(TGTACA) and d(CGATCG). 总被引:1,自引:0,他引:1
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4'-Deoxy-4'-iododoxorubicin, a halogenated anthracycline derivative, is an anticancer agent currently under Phase II clinical trials. In preclinical studies, it has demonstrated significantly reduced levels of cardiotoxicity compared to currently employed anthracyclines. It also has modified pharmacological properties resulting in an altered spectrum of experimental antitumor activity. The iodine atom at the 4' position of the sugar ring reduces the basicity and enhances the lipophilicity of this compound as compared to related anthracycline drugs. We report here single crystal X-ray diffraction studies of the complexes of 4'-deoxy-4'-iododoxorubicin with the hexanucleotide duplex sequences d(TGTACA) and d(CGATCG) at 1.6 and 1.5 A, respectively. The iodine substituent does not alter the geometry of intercalation as compared to previously solved anthracycline complexes, but appears to markedly affect the solvent environment of the structures. This could have consequences for the interaction of this drug with DNA and DNA binding proteins in cells. 相似文献
76.
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78.
付里叶变换红外光谱研究短杆菌肽A及其在脂双层中加Na~+前后的构象变化 总被引:1,自引:0,他引:1
用FTIR光谱研究了短杆菌肽A(GA)在固态,组装入DPPC脂持体,以及Na+结合状态的构象变化。固态时酰胺Ⅰ带峰位于163cm-1左右,且在1685cm-1处有一肩峰;酰胺Ⅱ带位于1530cm1左右,整个分子为反平行的β螺旋结构,为非通道构象。当GA组装入脂质体后,酰胺Ⅰ带仍位于1633cm-1,但1685cm-1的肩峰消失,酰胺Ⅱ带移至1550cm-1,分子间氢键转变为分子内氢键,二聚体从原来的双股螺旋变为头-头相连的单股螺旋,是GA的膜内通道构象。Na+的结合并不改变GA的骨架结构,但通过对酰胺Ⅰ带的拟合分析发现:加Na+后位于1644cm-1的无规结构含量减少10%,而有序结构增加,提示部分无视结构可能有一种“门”的作用:当无离子通过时为“关闭”状态,当与离子结合后即“打开”,这样通过构象的改变来实现其通道功能。 相似文献
79.
We have generated four xenogeneic rat antimouse IgG3 monoclonal antibodies recognizing at least three different antigenic determinants (epitopes) on BALB/c IgG3 molecules. These antibodies were used in solid-phase blocking radioimmunoassays for detection of the epitopes in sera of 40 inbred strains and 134 wild mice. These antibodies detect genetic polymorphism of IgG3 isotype among wild mice even though there is no polymorphism found among 40 inbred strains tested (except X-chromosome-linked immunodeficient CBA/N strain which lacks IgG3 molecules). An IgG3 variant was also isolated from hybridomas derived from Mus spretus.Abbreviations
Igh-C
immunoglobulin heavy chain constant region
- PVC
polyvinyl chloride
- RIA
radioimmune assay
- ELISA
enzyme linked immunosorbent assay 相似文献
80.
Human chromosomal assignments for 14 argininosuccinate synthetase pseudogenes: cloned DNAs as reagents for cytogenetic analysis 总被引:13,自引:6,他引:7
T S Su R L Nussbaum S Airhart D H Ledbetter T Mohandas W E O''Brien A L Beaudet 《American journal of human genetics》1984,36(5):954-964
There are multiple, processed, dispersed pseudogenes for human argininosuccinate synthetase. Chinese hamster X human somatic cell hybrids were used to map DNA fragment groups corresponding to the single expressed gene and 14 pseudogene loci. Each chromosomal assignment was confirmed using hybrids containing very few human chromosomes and/or by demonstrating monosomic or trisomic dosage in human cell lines with chromosomal abnormalities. Pseudogenes were mapped to chromosomes 2cen-p25, 3q12-qter, 4q21-qter, 5 (two loci), 6, 7, 9p13-q11, 9q11-q22, 11q, 12, Xp22-pter, Xq22-q26, and Ycen-q11. DNA fragments from the expressed gene were mapped to 9q34-qter in agreement with the previous assignment for enzyme activity. A high-frequency restriction fragment length polymorphism mapped to 9q11-q22. The analyses emphasized the feasibility of using chromosomally abnormal human cell lines for confirmation and regionalization of gene-mapping assignments made using somatic-cell hybrids. Conversely, cloned DNA probes, once mapped and characterized, can be very valuable for determining the chromosomal composition of interspecies hybrids and the dosage of loci in human cells. The argininosuccinate synthetase cDNA is a convenient reagent for dosage analysis of 15 human loci on 11 different chromosomes. Improved reagents could be designed that would simplify Southern blot patterns by eliminating overlapping DNA fragments and providing a single DNA fragment for each locus. 相似文献