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21.
Differential expression of two related amino acid transporters with differing substrate specificity in Arabidopsis thaliana 总被引:8,自引:3,他引:5
Marion Kwart Brigitte Hirner Sabine Hummel Wolf B. Frommer 《The Plant journal : for cell and molecular biology》1993,4(6):993-1002
A general amino acid permease cDNA ( AAP2 ) was isolated from Arabidopsis by complementation of a yeast mutant defective in citrulline uptake. Direct transport measurements in yeast show that the protein mediates uptake of l -[14 C]-citrulline and l -[14 C]-proline. Detailed analyses of the substrate specificity by competition studies demonstrate that all proteogenic amino acids are recognized by the carrier, including those that represent the major transport forms of reduced nitrogen in many species, i.e. glutamine, glutamate and asparagine. Thus, AAP2 is less selective as compared with AAP1 and transports basic amino acids such as histidine as shown by expression in a histidine transport-deficient yeast strain. The predicted polypeptide of 53 kDa is highly hydrophobic with 12 putative membrane-spanning regions and shows significant homologies to the Arabidopsis broad specificity permease AAP1, and a limited homology to bacterial branched chain amino acid transporters, but not to any other known proteins. Alterations in the charged residues as compared with AAP1 in four regions might be involved in the difference in selectivity towards basic amino acids. Both genes are highly expressed in developing pods indicating a role in supplying the developing seeds with reduced nitrogen. AAP2 is selectively expressed in the stem and might therefore play a role in xylem-to-phloem transfer of amino acids during seed filling. Furthermore in situ hybridization shows that both genes are expressed in the vascular system of cotyledons in developing seedlings. 相似文献
22.
We examined the effects of the conversion of tropical forest to pasture on soil organic matter (SOM) origin and quality along
a chronosequence of sites, including a primary forest and six pastures. Bulk soil samples received a physical size-fractionation
treatment to assess the contribution of each compartment to total SOM pool. Besides a general increase in total C and N stocks
along the chronosequence, we observed a reduction of the relative contribution of the coarser fractions to total soil C content,
and an increased concentration in the finer fractions. The origin of the C in each size fraction was established from measurements
of13C abundance. After 80 years about 93% of the C in the least humified fraction of the top 10 cm of soil was of pasture origin,
while in the most humified it was 82%. Chemical analyses indicated that the fine silt and coarse clay fractions contained
the most refractory carbon. 相似文献
23.
UV-induced allevation of lambda restriction in Escherichia coli K-12: kinetics of induction and specificity of this SOS function 总被引:11,自引:0,他引:11
Summary In UV-irradiated cells of Escherichia coli K-12 a partial release of the restriction of non-modified phage is observed when the cells are recA
+
lexA
+. We show here that the induction of this restriction allevation (RA) also depends on the recBC enzyme and that the expression of RA requires protein synthesis. Maximum expression was reached within 60 to 90 min after irradiation. Experiments are presented which show that upon UV-irradiation a signal is created which triggers the development of RA when protein synthesis is allowed. This signal decayed with a half-life of only a few minutes in cells treated with chloramphenicol. The decay kinetics were similar in uvr
+ and uvrA mutants. RA appeared to be specific for EcoK insofar as no allevation of restriction by EcoRI, EcoRII and EcoP1 occurred. During maximum expression of RA no gross reduction of the activities of the recBC enzyme (exonuclease V) and the restriction endonuclease EcoK was observed and no new DNA modifying activity appeared in the cells. Since, in fully expressed cells, up to 75% of the infecting DNA was converted to acid-soluble material within 20 min after infection we suggest that only a small specific fraction of infections may undergo RA. 相似文献
24.
Dr. Brigitte Krisch 《Cell and tissue research》1980,205(3):453-471
Summary The present ultrastructural results indicate that, in the rat, the vasopressin-synthesizing perikarya of the supraoptic nucleus (NSO) attain a certain degree of maturity earlier than those of the paraventricular nucleus (NPV). In the neonate rat, the stainability of the nuclear areas is very weak; in the perikarya of the NSO a few labeled granules can be found, whereas the perikarya of the NPV often display only a labeled Golgi area, the cytoplasm being devoid of granules. At the end of the first (NSO) and the second (NPV) postnatal weeks, the filling of the neurosecretory granules with vasopressin is inhomogeneous with irregular spots of reaction product distributed on the granules. This feature is less obvious during the following week and has nearly disappeared after the third and fourth postnatal weeks. Already in the neonate two types of vasopressin-positive fibers are observed in the median eminence, characterized by the different diameters of their granules and by their typical location in the internal and the external pericapillary contact zone. Especially in one and two week-old animals, in the internal zone of the median eminence and, to a lesser degree in the neural lobe, the immunocytochemical reaction product is deposited on an axonal tubular network. Judging from the presence of very few vasopressin-negative fibers in the neural lobe of the neonate, the development of the oxytocin system appears to be delayed. A characteristic relationship between pituicytes and the neurosecretory fibers can be observed during the first two postnatal weeks. After the third postnatal week the immunocytochemical features of the vasopressin system correspond approximately to that in adult rats.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/3) and Stiftung Volkswagenwerk 相似文献
25.
The measurement of the intrinsic alkaline Bohr effect of various human haemoglobins by isoelectric focusing 下载免费PDF全文
We have used isoelectric focusing to measure the differences between the pI values of various normal and mutant human haemoglobins when completely deoxygenated and when fully liganded with CO. It was assumed that the ΔpI(deox.–ox.) values might correspond quantitatively to the intrinsic alkaline Bohr effect, as most of the anionic cofactors of the haemoglobin molecule are `stripped' off during the electrophoretic process. In haemoglobins known to exhibit a normal Bohr coefficient (ΔlogP50/ΔpH) in solutions, the ΔpI(deox.–ox.) values are lower the higher their respective pI(ox.) values. This indicates that for any particular haemoglobin the ΔpI(deox.–ox.) value accounts for the difference in surface charges at the pH of its pI value. This was confirmed by measuring, by the direct-titration technique, the difference in pH of deoxy and fully liganded haemoglobin A0 (α2β2) solutions in conditions approximating those of the isoelectric focusing, i.e. at 5°C and very low concentration of KCl. The variation of the ΔpH(deox.–ox.) curve as a function of pH (ox.) was similar to the isoelectric-focusing curve relating the variation of ΔpI(deox.–ox.) versus pI(ox.) in various haemoglobins with Bohr factor identical with that of haemoglobin A0. In haemoglobin A0 the ΔpI(deox.–ox.) value is 0.17 pH unit, which corresponds to a difference of 1.20 positive charges between the oxy and deoxy states of the tetrameric haemoglobin. This value compares favourably with the values of the intrinsic Bohr effect estimated in back-titration experiments. The ΔpI(deox.–ox.) values of mutant or chemically modified haemoglobins carrying an abnormality at the N- or C-terminus of the α-chains are decreased by 30% compared with the ΔpI value measured in haemoglobin A0. When the C-terminus of the β-chains is altered, as in Hb Nancy (α2βTyr-145→Asp2), we observed a 70% decrease in the ΔpI value compared with that measured in haemoglobin A0. These values are in close agreement with the estimated respective roles of the two major Bohr groups, Val-1α and His-146β, at the origin of the intrinsic alkaline Bohr effect [Kilmartin, Fogg, Luzzana & Rossi-Bernardi (1973) J. Biol. Chem. 248, 7039–7043; Perutz, Kilmartin, Nishikura, Fogg, Butler & Rollema (1980) J. Mol. Biol. 138, 649–670]. In other mutant haemoglobins it is demonstrated also that the ΔpI(deox.–ox.) value may be decreased or even suppressed when the substitution affects residues involved in the stability of the tetramer. These results support the interpretation proposed by Perutz, Kilmartin, Nishikura, Fogg, Butler & Rollema [(1980), J. Mol. Biol. 138, 649–670] for the mechanism of the alkaline Bohr effect, and also indicate that the transition between the two quaternary configurations is a prerequisite for the full expression of the alkaline Bohr effect. 相似文献
26.
Summary Examination of serial semithin sections of rat kidney cortex and a subsequent electron microscopic study of selected areas revealed that the characteristic epithelium of the cortical part of the thick ascending limb of Henle extends for a varying distance beyond the macula densa. The transition from the relatively thin epithelium of the thick ascending limb at this site to the three -or even four-fold thicker epithelium of the convoluted part of the distal tubule is sharply defined and occurs without the interposition of an intermediate cell type.The position of the macula densa at the end but still clearly within the ascending limb of Henle's loop is functionally interpreted to guarantee the separation of the sensor point macula densa from disturbing influences which might arise from the secretory activity of the subsequent tubular portion.Investigations supported by the Deutsche Forschungsgemeinschaft. The skillful technical assistance of Mrs. Saliha Sabanovic is gratefully acknowledged 相似文献
27.
Further observations on cell wall morphogenesis and polysaccharide arrangement during plant growth 总被引:1,自引:1,他引:0
Summary The three-dimensional arrangement of the polysaccharide chains in cell walls was investigated, using ultracryotomy and cytochemistry, in order to test the validity of the previously postulated ordered fibril hypothesis and to analyze the characteristics of the primary wall morphogenesis.Both in mung bean hypocotyl (Phaseolus aureus) and pea root (Pisum sativum) cultured in defined conditions, cell to cell endogenous specificity is marked by differences in the numbers of layers, thickness, rhythm and direction of deposition. The occurrence of bow-shaped arrangements and of strata of orientation intermediate between the main crisscrossed multifibrillar layers suggests that the sequential changes of the morphogenetic activity of the cells is progressive. The twisted polysaccharide disposition evokes certain mesomorphic states; a part of the mechanism responsible for the wall arrangement may result from a self-assembly process as in the orientation of the molecules in a liquid cristal. This possibility finds experimental support in the fact that a three-dimensional association of the hemicellulose chains spontaneously appears when precipitated in acellular conditions.Polysaccharide removal associated with shadowing indicates that the ordered disposition within the wall is extensively altered by even a slight extraction. These data may invalidate diverse results which are generally brought forward to explain the wall organization during growth. 相似文献
28.
Brigitte Tréton Marie -Thérèse Le Dall H. Heslot 《Applied microbiology and biotechnology》1978,6(1):67-77
Summary We investigated the excretion of citric and isocitric acids in a strain of Saccharomycopsis lipolytica grown on either n-paraffins, glucose, or glycerol. These acids were excreted in the ratio of 67:33 on n-paraffins and roughly 92:8 on either glucose or glycerol. However, with all the carbon sources used, the relative amount of isocitric acid in the intracellular pool remained below 10%. The assimilation of citric and isocitric acids was prevented when glucose or glycerol were the carbon sources, but not when n-paraffins were used. Citric acid stopped isocitric acid assimilation. These phenomena of selective assimilation and/or uptake might explain the variations observed in the ratio of citric to isocitric acids excreted on different carbon sources. 相似文献
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