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The TATA-box binding protein (TBP) is one of the 4 DNA-binding proteins that has been shown to associate with the proximal
promoter region (−295) of the gene for bean seed storage protein phaseolin. The −295 promoter is essential for spatial and
temporal control of the phaseolin gene expression. We designed a pair of degenerated primers based on the highly conserved
sequence of the carboxyl-terminal domain of yeast TBP and used PCR to amplify the corresponding sequence from the bean cDNA.
By using the amplified fragment as a probe, we screened a cDNA library derived from poly A(+) RNA from developing bean seeds
and isolated 2 nearly full-length cDNA clones (813 and 826 bp long). The cDNAs encode 2 distinct isoforms of bean TBP, PV1
and PV2, each with an open reading frame of 200 amino acid residues. The 2 cDNA sequences share an 85.8% overall nucleotide
sequence identity, with the coding region showing a higher degree of identity (94.4%) than the 5′- and 3′-untranslated regions
(69%). The deduced amino acid sequence of the bean TBP isoforms differ in only 3 amino acid residues at positions 5, 9, and
16, all located in the amino-terminal region. The carboxyl-terminal domain of 180 amino acid residues shows a high degree
(>82%) of evolutionary sequence conservation with the TBP sequences from other eukaryotic species. This domain possesses the
3 highly conserved structural motifs, namely the 2 direct repeat sequences, a central basic region rich in basic amino acid
residues, and a region similar to the sigma factor of prokaryote. On the basis of this and other findings, we suggest that
higher plants in general may have at least 2 copies of TBP gene, presumably resulting from the global duplication of the genome.
Accession numbers AF015784 and AF015785 at the GenBank. 相似文献
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Vauclare Pierre Macherel David Douce Roland Bourguignon Jacques 《Plant molecular biology》1998,37(2):309-318
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Pierre Vauclare Richard Bligny Elisabeth Gout Valentine De Meuron François Widmer 《Planta》2010,231(6):1495-1504
The effects of dark-induced stress on the evolution of the soluble metabolites present in senescent soybean (Glycine max L.) nodules were analysed in vitro using 13C- and 31P-NMR spectroscopy. Sucrose and trehalose were the predominant soluble storage carbons. During dark-induced stress, a decline
in sugars and some key glycolytic metabolites was observed. Whereas 84% of the sucrose disappeared, only one-half of the trehalose
was utilised. This decline coincides with the depletion of Gln, Asn, Ala and with an accumulation of ureides, which reflect
a huge reduction of the N2 fixation. Concomitantly, phosphodiesters and compounds like P-choline, a good marker of membrane phospholipids hydrolysis
and cell autophagy, accumulated in the nodules. An autophagic process was confirmed by the decrease in cell fatty acid content.
In addition, a slight increase in unsaturated fatty acids (oleic and linoleic acids) was observed, probably as a response
to peroxidation reactions. Electron microscopy analysis revealed that, despite membranes dismantling, most of the bacteroids
seem to be structurally intact. Taken together, our results show that the carbohydrate starvation induced in soybean by dark
stress triggers a profound metabolic and structural rearrangement in the infected cells of soybean nodule which is representative
of symbiotic cessation. 相似文献
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Vauclare P Kopriva S Fell D Suter M Sticher L von Ballmoos P Krähenbühl U den Camp RO Brunold C 《The Plant journal : for cell and molecular biology》2002,31(6):729-740
The effect of externally applied L-cysteine and glutathione (GSH) on ATP sulphurylase and adenosine 5'-phosphosulphate reductase (APR), two key enzymes of assimilatory sulphate reduction, was examined in Arabidopsis thaliana root cultures. Addition of increasing L-cysteine to the nutrient solution increased internal cysteine, gamma-glutamylcysteine and GSH concentrations, and decreased APR mRNA, protein and extractable activity. An effect on APR could already be detected at 0.2 mm L-cysteine, whereas ATP sulphurylase was significantly affected only at 2 mm L-cysteine. APR mRNA, protein and activity were also decreased by GSH at 0.2 mm and higher concentrations. In the presence of L-buthionine-S, R-sulphoximine (BSO), an inhibitor of GSH synthesis, 0.2 mm L-cysteine had no effect on APR activity, indicating that GSH formed from cysteine was the regulating substance. Simultaneous addition of BSO and 0.5 mm GSH to the culture medium decreased APR mRNA, enzyme protein and activity. ATP sulphurylase activity was not affected by this treatment. Tracer experiments using (35)SO(4)(2-) in the presence of 0.5 mm L-cysteine or GSH showed that both thiols decreased sulphate uptake, APR activity and the flux of label into cysteine, GSH and protein, but had no effect on the activity of all other enzymes of assimilatory sulphate reduction and serine acetyltransferase. These results are consistent with the hypothesis that thiols regulate the flux through sulphate assimilation at the uptake and the APR step. Analysis of radioactive labelling indicates that the flux control coefficient of APR is more than 0.5 for the intracellular pathway of sulphate assimilation. This analysis also shows that the uptake of external sulphate is inhibited by GSH to a greater extent than the flux through the pathway, and that the flux control coefficient of APR for the pathway, including the transport step, is proportionately less, with a significant share of the control exerted by the transport step. 相似文献
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