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111.
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 下载免费PDF全文
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
112.
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
113.
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
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115.
水稻线粒体DNA雄性不育有关特异片段的克隆及序列分析 总被引:7,自引:0,他引:7
应用任意单引物聚合酶链反应技术,从水稻WA 型雄性不育系的线粒体DNA 中得到一个特异的扩增片段R2-630 WA。以该片段为探针进行Southern 杂交分析检测到在雄性不育胞质与正常可育胞质间存在的线粒体DNA 多态性。不育系珍汕97A 和其F1 杂种的杂交图谱相同。而保持系珍汕97B和恢复系明恢63 的杂交图谱一样。序列测定该片段全长629 bp,其碱基组成A+ T= 54.1% ,同源性比较结果显示, 该片段与1236 个已报道的植物基因(包括16 个水稻线粒体基因)序列的同源性均小于50% 。序列内含有一个长度为10 bp 的反向重复序列5-ACCATATGGT-3,位于262—272 区段。另外,其379—439 区段可编码一个含20个氨基酸残基的短肽。上述结果表明,R2-630 WA 片段确与水稻野败型雄性不育密切相关。推测反向重复序列5-ACCATATGGT-3在细胞质雄性不育性状形成中,可能起着重要作用 相似文献
116.
Mónica Suárez Margarita Martín Estrella Ferrer Amando Garrido-Pertierra 《Archives of microbiology》1995,164(1):70-77
Unlike the parent wild-type strain, theKlebsiella pneumoniae mutant strain MAO4 has a 4-HBA+ phenotype. The capacity of this mutant to take up and metabolize 4-hydroxybenzoate (4-HBA) relies on the expression of a
permease and an NADPH-linked monooxygenase (4-HBA-3-hydroxylase). Both enzymes are normally expressed at basal levels, and
only the presence of 4-HBA in the media enhances their activities. Strikingly, when theAcinetobacter calcoaceticus pobA gene encoding 4-hydroxybenzoate-3-hydroxylase was expressed in hydroxybenzoateK. pneumoniae wild-type, the bacteria were unable to grow on 4-HBA, suggesting that the main difference between the wild-type and the mutant
strain is the capability of the latter to take up 4-HBA. 4-HBA-3-hydroxylase was purified to homogeneity by affinity, gel-filtration,
and anion-exchange chromatography. The native enzyme, which appeared to be a dimer of identical subunits, had an apparent
molecular mass of 80 kDa and a pI of 4.6. Steady-state kinetics were analyzed; the initial velocity patterns were consistent
with a concerted substitution mechanism. The purified enzyme had 362 amino acid residues, and a tyrosine seemed to be involved
in substrate activation. 相似文献
117.
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 下载免费PDF全文
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
118.
Wei Wen Su 《Biotechnology Techniques》1995,9(4):259-264
Summary The effect of agitation and aeration on filtration of Anchusa officinalis culture in a stirred tank bioreactor integrated with an internal filter unit was investigated. Increases in suction head of the pump that drove the filtration process were measured at impeller speeds of 100 and 200 rpm. Surprisingly, suction head attained at 200 rpm was about 40% higher than at 100 rpm. Direct observation of the cake deposition process in the reactor using a dilute cell suspension revealed that the filter cake formed at 100 rpm was thicker, but less compact. Aeration at 0.4 vvm was shown to have little effect on the filtration rate, since the bulk fluid flow was dominated by the impeller hydrodynamics. The initial flux can be recovered by filter backwashing with compressed air at a flow rate of 0.6 vvm for a duration of 5 minutes. 相似文献
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