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111.
A plasmodial alpha-tubulin cDNA from Physarum polycephalum. Nucleotide sequence and comparative analysis 总被引:5,自引:0,他引:5
Günter Krämmer Monika Singhofer-Wowra Klaus Seedorf Melvyn Little Tim Schedl 《Journal of molecular biology》1985,183(4):633-638
As the first step towards correlating structure and function of tubulin in the slime mold Physarum polycephalum we have elucidated the nucleotide sequence of a cDNA that appears to code for all but the last 25 to 30 C-terminal amino acids of a plasmodial alpha-tubulin. Differences in amino acid sequence from those of other alpha-tubulins are distributed fairly evenly throughout the sequence, although a relatively extensive conserved region is found in position 396 to 426 near the C terminus. A small region in position 298 to 307 contains a cluster of amino acid residues unique to Physarum alpha-tubulin. The sequence is 70% homologous to two yeast alpha-tubulins and about 83% homologous to five animal alpha-tubulins. A comparison of the homologies of all the known alpha-tubulins indicates that a large decrease in the accepted point mutation rate has occurred during the evolution of the metazoa, suggesting a major functional specialization of microtubules. 相似文献
112.
Aprotic polar solvents inducing chromosomal malsegregation in yeast interfere with the assembly of porcine brain tubulin in vitro 总被引:2,自引:0,他引:2
A number of aprotic solvents which had previously been found to induce mitotic aneuploidy in yeast were tested for their effects on re-assembly of twice recycled tubulin from pig brain. Some of the solvents which were strong aneuploidy-inducing mutagens in yeast slowed down tubulin assembly in vitro at concentrations lower than those required for aneuploidy induction. Ethyl acetate, methyl acetate, diethyl ketone and acetonitrile fell into this category. Other strong aneuploidy-inducing agents like acetone and 2-methoxyethyl acetate accelerated tubulin assembly. Non-genetically active methyl isopropyl ketone and isopropyl acetate both accelerated assembly, whereas methyl n-propyl ketone and n-propyl acetate were weak inducers of aneuploidy and slowed down the rate and extent of assembly. Those chemicals which slowed down the assembly rate also reduced the extent of assembly. Most chemicals which accelerated assembly also led to an increased extent of assembly, with the exception of isopropyl acetate. At the higher concentrations, however, a maximum assembly rate was reached which was followed by a slow decline. Although a perfect correlation between effects on the induction of chromosomal malsegregation and the interference with tubulin assembly in vitro was not seen, the experiments with tubulin were carried out using this class of chemicals because some of them strongly induced mitotic aneuploidy under conditions which suggested tubulin to be the prime target. The lack of a perfect coincidence might be due to species differences between the porcine brain and the yeast spindle tubulin, or the test for aneuploidy induction may have been negative because the concentrations required for an effect on yeast tubulin may be greater than the general lethal toxicity limit. Bearing this reservation in mind, the results suggest that the yeast aneuploidy test has a considerable predictive value for mammalian mutagenicity. 相似文献
113.
The circadian movement of the lamina of primary leaves of Phaseolus coccineus L. is mediated by antagonistic changes in the length of the extensor and flexor cells of the laminar pulvinus. The cortex of the pulvinus is a concentric structure composed of hexagonal disc-like cells, arranged in longitudinal rows around the central stele. Observations with polarization optics indicate that the cellulose microfibrils are oriented in a hoop-like fashion in the longitudinal walls of the motor cells. This micellation is the structural basis of the anisotropic properties of the cells: tangential sections of the extensor and flexor placed in hypotonic mannitol solutions showed changes only in length. As a consequence a linear correlation between length and volume was found in these sections. Based on the relationship between the water potential (which is changed by different concentrations of mannitol) and the relative volume of the sections and on the osmotic pressure at 50% incipient plasmolysis, osmotic diagrams were constructed for extensor and flexor tissues (cut during night position of the pulvinus). The bulk moduli of extensibility,
, were estimated from these diagrams. Under physiological conditions the
values were rather low (in extensor tissue below 10 bar, in flexor tissue between 10 to 15 bar), indicating a high extensibility of the longitudinal walls of the motor cells. They are strongly dependent on the turgor pressure at the limits of the physiological pressure range.In well-watered plants, the water potentials of the extensor and flexor tissues were surprisingly low,-12 bar and-8 bar, respectively. This means that the cells in situ are by no means fully turgid. On the contrary, the cell volume in situ is similar to the volume at the point of incipient plasmolysis: the cell volumes of extensor and flexor cells in situ were only 1.01 times and 1.1 times larger, respectively, than at the point of incipient plasmolysis, whereas at full turgidity (cells in water) the corresponding factors were 1.8 and 1.5. It is suggested that the high elasticity of the longitudinal walls, the anisotropy of the cell walls, and the low water potential of the sections which is correlated with slightly stretched cell walls in situ, are favourable and effective for converting osmotic work in changes in length of the pulvinus cells, and thus for the up and down movement of the leaf.Symbols
volumetric elastic modulus
- i
instantaneous volumetric elastic modulus
- i
stationary volumetric elastic modulus
-
weight-averaged stationary bulk modulus of extensibility
- 0
osmotic pressure of the vacuole of a cell at the point of incipient plasmolysis
-
weight-averaged osmotic pressure of the vacuoles of the tissue at 50% incipient plasmolysis
-
water potential 相似文献
114.
Is the initiation of macronuclear DNA synthesis in Euplotes dependent on micronuclear functions? 总被引:1,自引:0,他引:1
To determine whether the micronucleus makes essential contributions during asexual reproduction, observations were made on cells of Euplotes octocarinatus from which the micronucleus had been removed with a micropipette. Most cells underwent one postenucleation division, then became arrested in macronuclear G1, slowed down in food uptake, developed macronuclear deformations, and finally died. Such cells could be rescued if a micronucleus was reimplanted before macronuclear deformations had developed. When provided with a new micronucleus, cells initiated macronuclear DNA synthesis about 12-16 h later. The data suggest that the micronucleus is involved in the control of the cell's transition from macronuclear G1 to S, and a model is proposed which postulates that in Euplotes macronuclear DNA synthesis is initiated when a micronucleus-encoded "initiator protein" has accumulated to a critical amount. 相似文献
115.
Penetration of C8 and C9 in the C5b-9 complex across the erythrocyte membrane into the cytoplasmic space 总被引:3,自引:0,他引:3
We have developed a technique in which transglutaminase is used to measure the penetration of terminal complement proteins across the erythrocyte membrane into the cytoplasmic space. Penetration of a given terminal complement protein into the cytoplasmic space was assessed by labeling the protein of interest with radioactive iodine, forming the complement channel using the labeled protein, adding transglutaminase to only one side of the membrane, and allowing the enzyme to cross-link the susceptible proteins on that side of the membrane. Cross-linking was assessed by measuring the increase in molecular weight of the appropriate molecule on sodium dodecyl sulfate gels under reducing conditions. The results of these experiments indicate that C8 and C9 are rapidly cross-linked to high molecular weight from either the interior or the exterior of the membrane. In order to determine whether the cross-linking mediated by enzyme on the interior was occurring from within the ghosts and not via enzyme that had leaked into the extracellular medium, experiments were performed with dimethylcasein in the extracellular medium. In the presence of this protein, cross-linking of C8 and C9 from outside was negligible. Hence, if cross-linking occurs when transglutaminase is trapped inside the ghosts, it cannot be due to leakage of enzyme, but must be attributable to cross-linking from the inside. The results show that C9 definitely penetrated across the membrane into the intracellular space. With respect to C8, statistical evaluation indicates that C8 probably penetrated into the intracellular space. 相似文献
116.
(±)-Abscisic acid (ABA) at 10-5 M was added to the transpiration stream of leaves of 16 species (C3 and C4, monocotyledons and dicotyledons). Stomatal responses followed one of three patterns: i) stomata that were wide and insensitive to CO2 initially, closed partially and became sensitive to CO2; ii) for stomata that were sensitive to CO2 before the application of ABA, the range of highest sensitivity to CO2 shifted from high to low intercellular partial pressures of CO2, for instance in leaves of Zea mays from 170–350 to 70–140 bar; iii) when stomata responded strongly to ABA, their conductance was reduced to a small fraction of the initial conductance, and sensitivity to CO2 was lost. The photosynthetic apparatus was affected by applications of ABA to various degrees, from no response at all (in agreement with several previous reports on the absence of effects of ABA on photosynthesis) through a temporary decrease of its activity to a lasting reduction. Saturation curves of photosynthesis with respect to the partial pressure of CO2 in the intercellular spaces indicated that application of ABA could produce three phenomena: i) a reduction of the initial slope of the saturation curve (which indicates a diminished carboxylation efficiency); ii) a reduction of the level of the CO2-saturated rate of assimilation (which indicates a reduction of the ribulose-1,5-bisphosphate regeneration capacity); and iii) an increase of the CO2 compensation point. Photosynthesis of isolated mesophyll cells was not affected by ABA treatments. Responses of the stomatal and photosynthetic apparatus were usually synchronous and often proportional to each other, with the result that the partial pressure of CO2 in the intercellular spaces frequently remained constant in spite of large changes in conductance and assimilation rate. Guard cells and the photosynthetic apparatus were able to recover from effects of ABA applications while the ABA supply continued. Recovery was usually partial, in the case of the photosynthetic apparatus occasionally complete. Abscisic acid did not cause stomatal closure or decreases in the rate of photosynthesis when it was applied during a phase of stomatal opening and induction of photosynthesis that followed a transition from darkness to light.Abbreviations and symbols
A
rate of CO2 assimilation
- ABA
(±)-abscisic acid
-
c
a
partial pressure of CO2 in the ambient air or in the gas supplied to the leaf chambers
-
c
i
partial pressure of CO2 in the intercellular spaces of a leaf
-
e
a
partial pressure of H2O in the air
-
g
conductance for water vapor
-
J
quantum flux
-
T
1
leaf temperature 相似文献
117.
Klaus Kuehn Udo Dunzendorfer W. F. Whitmore G. N. Schrauzer 《Biological trace element research》1985,8(4):237-250
Male Copenhagen rats with transplanted prostatic adeno-carcinoma were treated with different polyamine synthesis inhibitors, such as methylglyoxal-bis-guanylhydrazone (MGBG), erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) combined with 9-β-d-arabin-ofuranosyl-adenine (ARA-A), α-difluoromethyl-ornithine (DFMO), and some of their combinations. Levels of the essential trace elements—copper, zinc, magnesium, iron, selenium, and manganese —have been determined in blood, tumor, kidney, and liver of these animals and are discussed in terms of efficiency of the treatment. MGBG had the strongest effect on trace element levels in tissues of treated animals. MGBG combined with DFMO exhibited the highest antitumor activity of all treatment protocols. Selenium given as selenite with drinking water was used as an adjuvant with the most toxic combination, (ARA-A/EHNA, MGBG). Selenite reduced the toxicity of these therapeutic agents. 相似文献
118.
Andreas Leonhardt Estera Szwajcer Klaus Mosbach 《Applied microbiology and biotechnology》1985,21(3-4):162-166
Summary The relatively low solubility of oxygen in water presents a problem in particular when immobilized cells are used or enzymes are applied for oxygen dependent reactions. The other main purpose is the requirement of oxygen for the increase of biomass. In this investigation the usefulness of silicon emulsions as oxygen carriers is demonstrated.In case of l-amino acid oxidase activity of immobilized cells, an increase by a factor of four was found in the presence of silicon emulsions. Likewise, growth medium enriched with silicon emulsions showed a significantly increased growth of cells inside alginate beads compared to normal growth medium. 相似文献
119.
The effect of light on the metabolism of ammonia was studied by subjecting detached maize leaves to 150 or 1350 mol m–2 s–1 PAR during incubation with the leaf base in 2 mM 15NH4Cl. After up to 60 min, leaves were extracted. Ammonia, glutamine, glycine, serine, alanine, and aspartate were separated by isothermal distillation and ion exchange chromatography. 15N enrichments were analyzed by emission spectroscopy. The uptake of ammonium chloride did not influence CO2 assimilation (8.3 and 17.4 mol m–1 s–1 at 150 and 1350 mol m–2 s–1 PAR, respectively). Leaves kept at high light intensity contained more serine and less alanine than leaves from low light treatments. Within 1 h of incubation the enrichment of ammonia extracted from leaves rose to approximately 20% 15N. In the high light regime the amino acids contained up to 15% 15N, whereas in low light 15N enrichments were small (up to 6%). The kinetics of 15N incorporation indicated that NH3 was firstly assimilated into glutamine and then into glutamate. After 15 min 15N was also found in glycine, serine and alanine. At high light intensity nearly half of the 15N was incorporated in glycine. On the other hand, at low light intensity alanine was the predominant 15N sink. It is concluded that light influences ammonia assimilation at the glutamine synthetase reaction. 相似文献
120.
Christina Scharnhorst Hartmut Heinze Gabriele Meyer Waldemar Kolanus Klaus Bartsch Susanne Heinrichs Thomas Gudschun Margret Möller Frank Herzfeld 《Plant molecular biology》1985,4(4):241-245
Summary cDNA clones were isolated for a chloroplast protein, the mRNA of which is induced to maximum levels within 2–4 h after onset
of illumination in five day old, etiolated pea seedlings.
The cDNA library was constructed from poly(A)+-mRNA which was isolated from 4 h illuminated seedlings. The extremely short induction period of the early light induced protein(ELIP)-mRNA
established the basis of our screening procedure. Colony hybridization experiments were performed with32P-labelled cDNA probes, synthesized from RNA of seedlings which had been exposed to different programs of illumination. Plasmid
DNAs were isolated from colonies showing strong hybridization signals exclusively with cDNA corresponding to the 4 h-mRNA.
Hybrid released translation of preselected plasmids p 17/C2 and p17/C4 revealed a peptide of Mr 24 000. After posttranslational importin vitro, the processed product of Mr 17 000 appears in the chloroplast. Using these clones, the expression of the ELIP-mRNA was investigated by DOT-hybridization.
The ELIP-mRNA reaches maximum levels within 2–4 hours after onset of illumination. Our results correspond precisely to thein vivo characteristics and indicate positive identification of the sought clones. 相似文献