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Phytochrome-mediated regulation of two mRNAs, encoded by nuclei and chloroplasts of ribulose-1,5-bisphosphate carboxylase/oxygenase.

Etiolated pea seedlings (5 days old) were treated with red light for 5 min and grown for 2 days more in darkness. Effects of the red light on the mRNA levels of two subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) (ribulose-P2 carboxylase), rate of enzyme synthesis and accumulation of the enzyme were examined for the plants. Increases of the two mRNA levels, encoded in nuclei and chloroplasts were observed. Red light followed by far-red light treatment for 5 min inhibited the increases. An increase parallel with the mRNA levels was found for the rate of the enzyme synthesis and accumulation of the enzyme. These results indicate that the induction of two mRNA levels is mediated by phytochrome and causes the red-light-inducible, far-red-light-reversible increase of ribulose-P2 carboxylase. When white light was irradiated to induce the enzyme, the effects of red light given in darkness were reinforced. These results indicate that light-induction of ribulose-P2 carboxylase is mediated by phytochrome and controlled by the two mRNA levels.

Carboxy-Lyases↗

Fast Fluorescence Quenching from Isolated Guard Cell Chloroplasts of Vicia faba Is Induced by Blue Light and Not by Red Light.

Chlorophyll a fluorescence transients from isolated Vicia faba guard cell chloroplasts were used to probe the response of these organelles to light quality. Guard cell chloroplasts were isolated from protoplasts by passing them through a 10-mum nylon net. Intact chloroplasts were purified on a Percoll gradient. Chlorophyll a fluorescence transients induced by actinic red or blue light were measured with a fluorometer equipped with a measuring beam. Actinic red light induced a monophasic quenching, and transients induced by blue light showed biphasic kinetics having a slow and a fast component. The difference between the red and blue light-induced transients could be observed over a range of fluence rates tested (200-800 mumol m(-2) s(-1)). The threshold fluence rate of blue light for the induction of the fast component of quenching was 200 mumol m(-2) s(-1), but in the presence of saturating red light, fluence rates as low as 25 mumol m(-2) s(-1) induced the fast quenching. These results indicate that guard cell chloroplasts have a specific response to blue light.

Journal Article↗

Induction of atrial natriuretic factor and myosin light chain-2 gene expression in cultured ventricular myocytes by electrical stimulation of contraction.

While hormonal stimuli and mechanical stretch can induced cardiac-specific gene expression and in some cases cellular hypertrophy, the relationship between myocyte contraction frequency, gene expression, and myocyte growth has not been well characterized. In this study a new model system was developed in which cultures of neonatal rat ventricular myocytes were subjected to long term pacing of contractions with pulsatile electrical stimulation. Myocytes submitted to electrical stimulation for 3 days displayed dramatic increases in cellular size and myofibrillar organization, and a 5-10-fold increase in the expression of the cardiac genes atrial natriuretic factor and myosin light chain-2. Atrial natriuretic factor expression induced by electrical stimulation of contractions was inhibited by nifedipine or W7, indicating a dependence on calcium influx and calmodulin activity. Phosphoinositide hydrolysis and cAMP formation were not affected by electrical stimulation suggesting that gene induction occurred independently of the activation of protein kinases C or A above basal levels. These findings show that the cellular events associated with contraction, such as changes in cytoplasmic free calcium levels and/or cellular stretch, may serve as important determinants of myocyte growth and cardiac gene expression.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Mutagenic DNA repair in Escherichia coli. XVII. Effect of temperature-sensitive DnaE proteins on the induction of streptomycin-resistant mutations by UV light.

In contrast to the dnaE486 mutation, which is nearly 'dead-stop', dnaE1026 allows DNA synthesis for some time at restrictive temperatures. When bacteria carrying the dnaE486 or dnaE1026 temperature-sensitive mutations were incubated at restrictive temperature after exposure to UV light they showed little or no fixation of mutations as determined by loss of photoreversibility and the mutation frequency fell progressively. These results confirm a role for DnaE protein (the alpha-subunit of DNA polymerase III holoenzyme) in UV mutagenesis. A derivative of dnaE1026 carrying the umuC122 allele which blocks normal UV mutagenesis showed induction of mutations when photoreversing light was given to UV-irradiated bacteria after a period of incubation at either permissive or restrictive temperature. The defective DnaE1026 protein is therefore able to carry out the misincorporations which have been postulated to be the first step in the mutagenic process. Its inability to give rise to mutations in umu+ bacteria may therefore be attributed to its inability to participate in a later step, perhaps because it is unable to interact with the UmuD, C gene products. In contrast, UV-irradiated dnaE486 umuC122 bacteria did not show mutagenesis when incubated at restrictive temperature before photoreversal, suggesting that the altered DnaE486 protein was not able to carry out the postulated misincorporation step at 43 degrees C. DNA polymerase III alpha-subunit therefore appears to be required for both the misincorporation and bypass steps in the two-step model for UV mutagenesis.

DNA Repair↗

The interaction of respiration and photosynthesis in induction of nitrate reductase activity.

The respiration and photosynthesis requirement for induction and maintenance of nitrate reductase activity was determined on leaves of Hordeum vulgare L. In this induction, glucose substituted for light in both dark-grown and carbohydrate-depleted green leaves. Oxygen appeared to be required for induction in all cases studied. In light and under N(2), 3-(3,4-dichlorophenyl)-1,1-dimethylurea completely inhibited induction, presumably by inhibiting the production of O(2), Hence, under N(2) the leaves appeared to utilize both the O(2) produced by photosynthesis and the CO(2) produced by respiration. CO(2) fixation can then produce both photosynthate to drive the induction and terminal electron acceptors to allow photosynthetic electron flow. This possibility was further suggested by the observation that CO(2) was an absolute requirement for induction in carbohydrate-depleted barley leaves. Results obtained with respiratory inhibitors also indicated that respiration drove the induction of nitrate reductase.Exogenously supplied glucose also substantially slowed the loss of nitrate reductase that occurred when barley leaves were placed in darkness. It is presumed that glucose allowed the synthetic or activation phase of the induction to proceed more rapidly. Our results support the hypothesis that one of the main effects of light may be to supply photosynthate to support respiration, which then drives the induction process.

Journal Article↗

Photic and circadian regulation of c-fos gene expression in the hamster suprachiasmatic nucleus.

Photic information entrains a circadian pacemaker located in the suprachiasmatic nucleus (SCN) of the mammalian hypothalamus to environmental light/dark cycles. To determine whether light regulates c-fos gene expression in the SCN, we have measured c-fos mRNA levels in the SCN of the golden hamster. We report that, during the subjective night, light causes a rapid increase in levels of c-fos mRNA in the SCN. Light pulses of 5 min duration are sufficient to induce c-fos mRNA, and the highest mRNA levels occur 30 min following the onset of light. The minimum level of illumination required to induce an increase in c-fos mRNA is indistinguishable from the minimum irradiance that produces a phase shift in the hamster's circadian rhythm of activity. In addition, the induction of c-fos mRNA in the SCN by light is itself under circadian regulation. Light induction of c-fos mRNA occurs only during the subjective night, at circadian times when photic phase shifting of activity occurs. Taken together, these data suggest that c-fos may be a molecular component of the photic pathway for entrainment of mammalian circadian rhythms.

Animals↗

Regulatory factors controlling photosynthetic reaction center and light-harvesting gene expression in Rhodobacter capsulatus.

Most species of photosynthetic bacteria synthesize their photosynthetic apparatus only under conditions of reduced oxygen tension. To a large extent, this phenomenon is dependent upon anaerobic induction of photosynthesis gene expression. Here we report an example of a regulatory gene, regA, that is involved in transactivating anaerobic expression of the photosynthetic apparatus. We show that RegA is itself responsible for differential induction of light-harvesting and reaction center gene expression relative to operons for photopigment biosynthesis. Surprisingly, strains disrupted for regA were found to retain normal photosynthetic growth capabilities under high light intensities. We further show that photosynthetic growth in the absence of transactivating structural gene expression is a consequence of the superoperonal organization of the photosynthetic gene cluster.

Amino Acid Sequence↗

High light-induced switch from C(3)-photosynthesis to Crassulacean acid metabolism is mediated by UV-A/blue light.

The high light-induced switch in Clusia minor from C(3)-photosynthesis to Crassulacean acid metabolism (CAM) is fast (within a few days) and reversible. Although this C(3)/CAM transition has been studied intensively, the nature of the photoreceptor at the beginning of the CAM-induction signal chain is still unknown. Using optical filters that only transmit selected wavelengths, the CAM light induction of single leaves was tested. As controls the opposite leaf of the same leaf pair was studied in which CAM was induced by high unfiltered radiation (c. 2100 micromol m(-2) s(-1)). To evaluate the C(3)-photosynthesis/CAM transition, nocturnal CO(2) uptake, daytime stomatal closure and organic acid levels were monitored. Light at wavelengths longer than 530 nm was not effective for the induction of the C(3)/CAM switch in C. minor. In this case CAM was present in the control leaf while the opposite leaf continued performing C(3)-photosynthesis, indicating that CAM induction triggered by high light conditions is wavelength-dependent and a leaf internal process. Leaves subjected to wavelengths in the range of 345-530 nm performed nocturnal CO(2) uptake, (partial) stomatal closure during the day (CAM-phase III), and decarboxylation of citric acid within the first 2 d after the switch to high light conditions. Based on these experiments and evidence from the literature, it is suggested that a UV-A/blue light receptor mediates the light-induced C(3)-photosynthesis/CAM switch in C. minor.

Adaptation, Physiological↗

Evaluation of apoptotic cells induced by ultraviolet light B radiation in epidermal sheets stained by the TUNEL technique.

Two major components of epidermal cells, keratinocytes and Langerhans cells, are injured by ultraviolet light B radiation, resulting in sunburn cell (apoptotic cell) formation, impaired function, and a reduced number of Langerhans cells. Quantitative analysis of Langerhans cell damage is usually performed using epidermal sheets, whereas that of keratinocytes has been performed by counting the number of sunburn cells in vertical tissue sections. In this study we assessed the influences of ultraviolet light B radiation on epidermal cells by apoptotic cell formation, using murine epidermal sheets stained by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling technique. Ten to 75 mJ per cm2 of ultraviolet light B radiation induced apoptotic cells in abdominal skin of C3H mice. The cells were induced in 6 h after 50 mJ per cm2 of ultraviolet light B irradiation with the peak in number in 24 h, 18.8 +/- 5.0 per mm2 and 97.7 +/- 7.4 per mm2, respectively. One week later, the apoptotic cells were not visualized. As C3H/He, BALB/C, and C57BL/6 mice showed almost the same frequency of apoptosis in epidermal sheets from 50 mJ per cm2 ultraviolet light B-irradiated skin, the induction of the cells by ultraviolet light B radiation did not depend on the genetic trait of the mouse. Xeroderma pigmentosum type A gene-deficient mice, however, showed a greater induction of apoptotic cells (216.9 +/- 25.2 per mm2) by ultraviolet light B radiation than xeroderma pigmentosum type A wild-type mice (89.5 +/- 13.6 per mm2) and conventional mice. Pretreatment with a SPF 60 sunscreen agent was quite effective in reducing the induction of apoptotic cells. Using confocal laser scanning microscopy and double staining, 1.5 +/- 2.7% of apoptotic cells were Ia-positive cells in 24 h after 50 mJ per cm2 of ultraviolet light B radiation. Apoptotic Ia-positive cells were not observed 48 h after the radiation. On the other hand, no apoptotic dendritic epidermal T cells were observed in up to 75 mJ per cm2 of ultraviolet light B radiated skin. Thus, nearly all apoptotic cells were keratinocytes, and Langerhans cells and dendritic epidermal T cells appeared resistant to ultraviolet light B-induced apoptosis. Compared with the assessment in vertical tissue sections, the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling technique with epidermal sheets appeared to be a more physiologically relevant method for quantitative evaluation of apoptotic epidermal cells induced by ultraviolet light B radiation.

Animals↗

Time course and quantum efficiency of photosynthesis in Chlorella.

1. Though the quantum yield remains constant for different samples of the same culture despite great changes in respiration due to dark adaptation, the quantum requirement for different cultures varies from 6.1 to 13.5 quanta per molecule of oxygen evolved (q/m). 2. This variation from one culture to another appears to depend upon chlorophyll concentration, though other paralleling factors cannot be ruled out. 3. Both chlorophyll concentration and quantum requirement show a random distribution. A statistical median for 50 cultures and 99 determinations gives q/m = 8.5 with a systematic uncertainty of perhaps 10 per cent. Since the variations are real, the median is regarded as less important than the lower limit approached (about q/m = 6). 4. Dark adaptation under aerobic conditions produces an initial photosynthetic rate of nearly zero. The immediate rise to steady state is somewhat logarithmic in character and may require over 3 minutes. 5. In intermittent light (of periods from 1 to 6 minutes) the induction observed in subsequent light periods starts from a finite initial rate and occupies a shorter time, often as little as 30 seconds. 6. The theoretical importance of aerobic induction is discussed. A chlorophyll cycle of two photochemical steps is found to satisfy most of the observed characteristics and to be compatible with an efficiency independent of intensity.

Chlorella↗

Gravitational response of the slime mold Physarum.

The acellular slime mold Physarum polycephalum is used as a model system to investigate the graviresponse of single cells which possess no receptors specialized for the perception of gravity. To obtain insights into the gravity-signal transduction mechanism the light response of the cell is used: Macroplasmodia of the slime mold show clear geo- and phototaxes. Gravity increases and white light decreases transiently the contraction frequency of plasmodial strands whereby both responses follow the same time pattern. Since mitochondria play a major role in changing the contraction rhythm in response to light and gravity stimuli, the simultaneous and subsequent inductions of the opposing light and gravity responses and their mutual influences on one another were investigated. The experiments were performed in weightlessness (0 g)--simulated on the fast-rotating clinostat as well as in actual weightlessness during the IML-1 Space Shuttle mission. The results indicate that mitochondria (chondriome) are part of the acceleration-stimulus reaction chain in Physarum. Two models for a direct gravireceptor mechanism are discussed.

Animals↗

Simultaneous S-cone contrast.

Chromatic induction is the change in appearance of one light caused by a second, nearby light. We measured chromatic induction in a central test viewed within an inducing field that was varied in only short-wavelength-sensitive (S) cone stimulation. The observer matched the appearance of the central test by adjusting the chromaticity of a haploscopically presented comparison field, seen by the other eye on a dark background. When the central test weakly stimulated S cones, the S-cone level in the surround caused little change in the color appearance of the test. When the central test substantially stimulated S cones, on the other hand, the appearance of the center showed S-cone contrast: raising the level of S in the surround reduced the level of S set to match the central test. Further, a surround that weakly stimulated S cones raised the matching S-cone level above that required without a surround (dark-adapted condition). These results cannot be explained by S-cone sensitivity loss or by a two-process model of adaptation. A cortical mechanism is proposed to mediate S-cone antagonism.

Color Perception↗

Development and characterization of a whole-cell bioluminescent sensor for bioavailable middle-chain alkanes in contaminated groundwater samples.

A microbial whole-cell biosensor was developed, and its potential to measure water-dissolved concentrations of middle-chain-length alkanes and some related compounds by bioluminescence was characterized. The biosensor strain Escherichia coli DH5 alpha(pGEc74, pJAMA7) carried the regulatory gene alkS from Pseudomonas oleovorans and a transcriptional fusion of PalkB from the same strain with the promoterless luciferase luxAB genes from Vibrio harveyi on two separately introduced plasmids. In standardized assays, the biosensor cells were readily inducible with octane, a typical inducer of the alk system. Light emission after induction periods of more than 15 min correlated well with octane concentration. In well-defined aqueous samples, there was a linear relationship between light output and octane concentrations between 24 and 100 nM. The biosensor responded to middle-chain-length alkanes but not to alicyclic or aromatic compounds. In order to test its applicability for analyzing environmentally relevant samples, the biosensor was used to detect the bioavailable concentration of alkanes in heating oil-contaminated groundwater samples. By the extrapolation of calibrated light output data to low octane concentrations with a hyperbolic function, a total inducer concentration of about 3 nM in octane equivalents was estimated. The whole-cell biosensor tended to underestimate the alkane concentration in the groundwater samples by about 25%, possibly because of the presence of unknown inhibitors. This was corrected for by spiking the samples with a known amount of an octane standard. Biosensor measurements of alkane concentrations were further verified by comparing them with the results of chemical analyses.

Aldehydes↗

Conidiation in Penicillium cyclopium is induced by conidiogenone, an endogenous diterpene.

The filamentous fungus Penicillium cyclopium conidiates in the presence of calcium ions in submerged culture without nutrient limitation according to a precisely timed program. Conidiation could be prematurely induced in a nutritionally sufficient medium which had previously supported growth, suggesting that a metabolite which influenced the process was produced. A diterpenoid with conidiation-inducing activity, conidiogenone, was purified from the culture medium, along with conidiogenol, a putative derivative with delayed activity. Contrary to previous thought, the presence of calcium was demonstrated to only decrease the threshold concentration of conidiogenone required for the induction to proceed. In light of these results, a mechanism of conidiation induction is presented according to which the mycelium produces a conidiation inducer (conidiogenone) that accumulates extracellularly. When a threshold concentration is reached, induction likely takes place by interaction with a specific cellular receptor. The results indicate that conidiogenone is both sufficient and necessary to induce conidiation.

Calcium↗

LIGHT, a new TNF superfamily member, is essential for memory T helper cell-mediated activation of dendritic cells.

LIGHT is a recently identified member of the TNF superfamily that is up-regulated on activated T cells and can cooperate with CD40L to condition DC for the priming of CTL. In this report, we show that an intracellular pool of LIGHT is found selectively in some CD45RA-CD27+CD4+ "central" memory T cells and to a greater extent in some CD45RA-CD27-CD4+ effector memory T cells. LIGHT, like CD40L, is rapidly up-regulated on memory CD4+ T cells upon activation. Unlike CD40L, LIGHT up-regulation on naive T cells is delayed. Stimulation of DC with each subset of memory T cells in the presence of superantigen revealed that CD40L and LIGHT are required for optimal secretion of IL-12. Moreover, effector memory T cells, which can interact with DC in peripheral tissues, contribute to CCR7 induction on DC. LIGHT and CD40L can also regulate IL-12 production by CCR7+ DC capable of migration to lymph nodes. These data suggest that both LIGHT and CD40L may be involved in the maintenance or reactivation of secondary Th1 responses.

CD4-Positive T-Lymphocytes↗

The promoter of a pine photosynthetic gene allows expression of a beta-glucuronidase reporter gene in transgenic rice plants in a light-independent but tissue-specific manner.

In angiosperms, the expression of the cab gene that encodes the chlorophyll a/b-binding protein of PSII is light-regulated. However, the pine cab gene is expressed in a light-independent but cell-type-specific manner. In the present study, the cab-6 promoter (1.7 kbp) from pine was fused to a beta-glucuronidase (GUS) reporter gene and the chimeric gene was introduced into rice protoplasts by electroporation. The GUS expression was studied in the resultant transgenic rice plants. Expression of GUS at a substantial level was confirmed in primary leaves of dark-germinated rice seedlings, and no obvious effect of light on the GUS activity was observed. The expression of GUS was restricted to photosynthetic tissues. The pine cab-6 promoter is, thus, sufficient for induction of light-independent but cell-type-specific expression in cells of a monocot, as is the case in the original pine cells.

Base Sequence↗

[Ultraviolet irradiation of Saccharomyces cerevisia: variations during meiosis in survival and in the induction of the cytoplasmic "petite" mutation].

Cyclic variations in sensitivity to killing and cytoplasmic "petite' (p-) induction by ultraviolet light occur during the meiosis of Saccharomyces cerevisiae. Maximal sensitivity to killing coincides with the period of meiotic nuclear DNA synthesis. Cyclic fluctuations in p- induction could not be correlated with known meiotic events and the pattern could vary temporarily from batch to batch. A dark liquid holding of irradiated cells aided the repair of lethal lesions but on the other hand an enhancement of "petite" induction was observed at all meiotic stages.

Cytoplasm↗