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We have studied by electron microscopy the size and morphology of the complexes obtained with different DNAs (between 500 and 5243 base pairs long) and four different proteins: sea urchin histone H1; sea cucumber histone ?0, chicken erythrocyte histone H5, and clupeine. Surprisingly, the type of protein used has only a marginal influence on the complexes formed. The molecular weight and topology of DNA do not show any influence. The size of the complexes depends strongly on the ratio of positive to negative charges and also on the ionic conditions. Our studies have been mainly carried out at a ratio of 0.4. Under these conditions the average thickness of rods and toroids observed varies between 165 Å at 1.5 mM salt to 290 Å at 100 mM salt, with minor variations around these values depending on the type of DNA and protein used. We conclude that the formation of DNA condensates is mainly determined by a balance of electrostatic and intermolecular forces, the influence of specific interactions is only marginal. This conclusion seems to apply not only to the complexes described here, but also to chromatin fibers and to DNA condensed by low molecular weight counterions and other compounds (polyamines, inorganic ions, ethanol, etc.). © 1994 John Wiley & Sons, Inc. 相似文献
85.
Summary The types and amount of plasma membrane proteins synthesized during cell elongation in response to auxin (2,4-dichlorophenoxyacetic acid) treatment were investigated. Auxin-treated and control soybean (Glycine max L.) hypocotyl segments were incubated with [35S]methionine for various times, ranging from 0.5 to 18 h, prior to isolation of plasma membrane by aqueous two-phase partitioning. Protein accumulated in the plasma membrane after auxin treatment. Despite this accumulation, the protein incorporation rate, estimated by the amount of label in the plasma membrane following a 0.5 h [35S]methionine pulse, was unaffected by auxin treatment at both 0.5 and 18 h of treatment. Protein apparently accumulated by a mechanism distinct from enhanced incorporation. The plasma membrane proteins synthesized by elongating segments differed from controls at 18 h, as evidenced by the pattern of fluorographs following a 0.5 h radiolabelling. However, auxin treatment did not alter the 2-D gel pattern of the polypeptides detectable by silver stain.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IEF
isoelectric focusing
- PM
plasma membrane
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
86.
87.
Non-photochemical fluorescence quenching and the diadinoxanthin cycle in a marine diatom 总被引:7,自引:0,他引:7
Miguel Olaizola Julie La Roche Zbigniew Kolber Paul G. Falkowski 《Photosynthesis research》1994,41(2):357-370
The diadinoxanthin cycle (DD-cycle) in chromophyte algae involves the interconversion of two carotenoids, diadinoxanthin (DD) and diatoxanthin (DT). We investigated the kinetics of light-induced DD-cycling in the marine diatom Phaeodactylum tricornutum and its role in dissipating excess excitation energy in PS II. Within 15 min following an increase in irradiance, DT increased and was accompanied by a stoichiometric decrease in DD. This reaction was completely blocked by dithiothreitol (DTT). A second, time-dependent, increase in DT was detected 20 min after the light shift without a concomitant decrease in DD. DT accumulation from both processes was correlated with increases in non-photochemical quenching of chlorophyll fluorescence. Stern-Volmer analyses suggests that changes in non-photochemical quenching resulted from changes in thermal dissipation in the PS II antenna and in the reaction center. The increase in non-photochemical quenching was correlated with a small decrease in the effective absorption cross section of PS II. Model calculations suggest however that the changes in cross section are not sufficiently large to significantly reduce multiple excitation of the reaction center within the turnover time of steady-state photosynthetic electron transport at light saturation. In DTT poisoned cells, the change in non-photochemical quenching appears to result from energy dissipation in the reaction center and was associated with decreased photochemical efficiency. D1 protein degradation was slightly higher in samples poisoned with DTT than in control samples. These results suggest that while DD-cycling may dynamically alter the photosynthesis-irradiance response curve, it offers limited protection against photodamage of PS II reaction centers at irradiance levels sufficient to saturate steady-state photosynthesis.Abbreviations CAP
chloramphenicol
- D1
PS II reaction center protein
- DD
diadinoxanthin
- DD
cycle-diadinoxanthin cycle
- DT
diatoxanthin
- DTT
dithiothreitol
- FCP
fucoxanthin chlorophyll a-c protein
- Fm
maximum fluorescence yield in the dark-adapted state
- Fo
minimum fluorescence yield in the dark-adapted state
- Fm and Fo
maximum and minimum fluorescence yields respectively in some light adapted state
- Fv
maximum variable fluorescence yield in the dark-adapted state
- Ik
Irradiance at the intercept of the initial slope of the photosynthesis-irradiance curve and the maximum photosynthetic rate
- kD
first order rate constant for nonradiative de-excitation of excitions in the PS II antenna
- kd
first order rate constant for non-radiative de-excitation of excitons in the PS II reaction center
- kF
first order rate constant for fluorescence
- kT
first order rate constant for exciton transfer to the reaction center
- kt
first order rate constant for exciton transfer from the reaction center to the antenna
- Rubisco
ribulose bisphosphate carboxylase
- SVm
Stern-Volmer quenching coefficient of the maximum fluorescence yield
- SVo
Stern-Volmer quenching coefficient of the miniximum fluorescence yield
- PS II
apparent absorption cross-section of PS II
- arr
average interval between exciton arrival to the PS II reaction center (ms)
- rem
average interval between electron turnover during photosynthesis in the PS II reaction center (ms)
- d
the probability that an exciton is non-radiatively dissipated in the reaction center
- T
the probability that an exciton in the antenna is transferred to the reaction center
- t
the probability that an exciton is transferred back from the reaction center to the antenna 相似文献
88.
89.
José Antonio Jarillo Juan Capel Antonio Leyva José Miguel Martínez-Zapater Julio Salinas 《Plant molecular biology》1994,25(4):693-704
We have isolated two Rare Cold-Inducible (RCI1 and RCI2) cDNAs by screening a cDNA library prepared from cold-acclimated etiolated seedlings of Arabidopsis thaliana with a subtracted probe. RNA-blot hybridizations revealed that the expression of both RCI1 and RCI2 genes is induced by low temperature independently of the plant organ or the developmental stage considered. However, RCI1 mRNA accumulates faster and at higher levels than the RCI2 one indicating that these genes have differential responsiveness to cold stress. Additionally, when plants are returned to room temperature, RCI1 mRNA decreases faster than RCI2. In contrast to most of the cold-inducible plant genes characterized, the expression of RCI1 and RCI2 is not induced by ABA or water stress. The nucleotide sequences of RCI1 and RCI2 cDNAs predict two acidic polypeptides of 255 and 251 amino acids with molecular weights of 29 and 28 kDa respectively. The alignment of these polypeptides indicates that they have 181 identical amino acids suggesting that the corresponding genes have a common origin. Sequence comparisons reveal no similarities between the RCI proteins and any other cold-regulated plant protein so far described. Instead, they demonstrate that the RCI proteins are highly homologous to a family of proteins, known as 14-3-3 proteins, which are thought to be involved in the regulation of multifunctional protein kinases. 相似文献