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At least 451 records · Page 25Linked to original sources

Light-Stimulated Apical Hook Opening in Wild-Type Arabidopsis thaliana Seedlings.

Apical hook opening and cotyledon unfolding are characteristic responses that occur during deetiolation of dicotyledonous seedlings. Light-stimulated apical hook opening and cotyledon unfolding in etiolated Arabidopsis thaliana seedlings appears to involve the activities of multiple photosensory systems. Red, far-red, and blue light are all effective in stimulating these responses in Arabidopsis. Stimulation of hook opening by red light and low fluence blue light is inductive, far-red reversible, and exhibits reciprocity, as is characteristic of many low fluence-dependent phytochrome-mediated responses. Far-red and high-fluence blue light appear to stimulate hook opening and cotyledon unfolding through high-irradiance-response systems during long-term light treatments. Although a phytochrome high-irradiance-response system presumably mediates the responses in far-red light, the responses to high-fluence blue light may be mediated by a blue light-specific photosensory system.

Journal Article↗

Genotoxic profile of inhibitors of topoisomerases I (camptothecin) and II (etoposide) in a mitotic recombination and sex-chromosome loss somatic eye assay of Drosophila melanogaster.

Genotoxic carcinogens which interact with DNA may produce double-strand breaks as normal intermediates of homologous mitotic recombination, and may give rise to structural chromosome aberrations and inter-chromosomal deletion-recombination. The genotoxic profile of two inhibitors of DNA topoisomerases were evaluated using an in vivo somatic w/w+ eye assay of Drosophila melanogaster for the detection of loss of heterozygosity (LOH) by homologous mitotic recombination, intra-chromosomal recombination and structural chromosomal aberrations. We studied camptothecin (CPT) as a topoisomerase-I-interactive agent and etoposide (ETOP) as a topoisomerase II inhibitor. These drugs act by stabilizing a ternary complex consisting of topoisomerases covalently linked to DNA at single-strand or at double-strand breaks, thereby preventing the relegation step of the breakage/rejoining reaction mediated by the enzyme. The genotoxic profiles were determined from the appearance of eye tissue in adult flies, in which LOH and expression of the reporter gene white produced light clones. The results demonstrated that both compounds were significantly genotoxic, with CPT being more effective than ETOP. Inter-chromosomal mitotic recombination was the major mechanism responsible for the induction of light spots by both compounds in XX females. Loss of the ring X chromosome (rX), was significantly enhanced by CPT, and this topoisomerase blocker also produced intra-chromosomal recombination (XY males).

Animals↗

Coordinate regulation of energy transduction modules in Halobacterium sp. analyzed by a global systems approach.

The extremely halophilic archaeon Halobacterium NRC-1 can switch from aerobic energy production (energy from organic compounds) to anaerobic phototrophy (energy from light) by induction of purple membrane biogenesis. The purple membrane is made up of multiple copies of a 1:1 complex of bacterioopsin (Bop) and retinal called bacteriorhodopsin that functions as a light-driven proton pump. A light- and redox-sensing transcription regulator, Bat, regulates critical genes encoding the biogenesis of the purple membrane. To better understand the regulatory network underlying this physiological state, we report a systems approach using global mRNA and protein analyses of four strains of Halobacterium sp.: the wild-type, NRC-1; and three genetically perturbed strains: S9 (bat+), a purple membrane overproducer, and two purple membrane deficient strains, SD23 (a bop knockout) and SD20 (a bat knockout). The integrated DNA microarray and proteomic data reveal the coordinated coregulation of several interconnected biochemical pathways for phototrophy: isoprenoid synthesis, carotenoid synthesis, and bacteriorhodopsin assembly. In phototrophy, the second major biomodule for ATP production, arginine fermentation, is repressed. The primary systems level insight provided by this study is that two major energy production pathways in Halobacterium sp., phototrophy and arginine fermentation, are inversely regulated, presumably to achieve a balance in ATP production under anaerobic conditions.

Adenosine Triphosphate↗

Role of ubiquitin carboxy terminal hydrolase-L1 in neural cell apoptosis induced by ischemic retinal injury in vivo.

Ubiquitin is thought to be a stress protein that plays an important role in protecting cells under stress conditions; however, its precise role is unclear. Ubiquitin expression level is controlled by the balance of ubiquitinating and deubiquitinating enzymes. To investigate the function of deubiquitinating enzymes on ischemia-induced neural cell apoptosis in vivo, we analyzed gracile axonal dystrophy (gad) mice with an exon deletion for ubiquitin carboxy terminal hydrolase-L1 (UCH-L1), a neuron-specific deubiquitinating enzyme. In wild-type mouse retina, light stimuli and ischemic retinal injury induced strong ubiquitin expression in the inner retina, and its expression pattern was similar to that of UCH-L1. On the other hand, gad mice showed reduced ubiquitin induction after light stimuli and ischemia, whereas expression levels of antiapoptotic (Bcl-2 and XIAP) and prosurvival (brain-derived neurotrophic factor) proteins that are normally degraded by an ubiquitin-proteasome pathway were significantly higher. Consistently, ischemia-induced caspase activity and neural cell apoptosis were suppressed approximately 70% in gad mice. These results demonstrate that UCH-L1 is involved in ubiquitin expression after stress stimuli, but excessive ubiquitin induction following ischemic injury may rather lead to neural cell apoptosis in vivo.

Animals↗

[Photomorphogenesis].

The normal development of the higher plant occurs only in light (photomorphogenesis). The effect of light is due to intracellular development of a morphogenetically active effector molecule (Pfr, a chromoprotein). The point in question is, by which pathways the homeostasis of development (the course of development directed by endogenic factors) and the environmental factor "light" may act together in photomorphogenesis in order to accomplish the normal development of plants. Physiological and molecular analysis of photomorphogenesis is to contribute basically to the problem of surveying the pathways of gens and environmental influences in determining the characteristics of higher organisms, man included. The logical order of the phenomena is shown to have the precedence of an attempt for molecular analysis of photomorphogenesis. A result of the logical order is the insight that all the phenomena induced by Pfr present the spatial and temporal pattern of "primary differentiation". This primary differentiation (= formation of the specific competence) does not depend on light (developmental homeostasis). The photomorphogenesis (= development through the presence of Pfr of the pattern due to the primary differentiation), in accordance with the present knowledge is to be attributed to differential enzymatic induction and repression. This opinion is confirmed by examples. The correlation between the enzymatic activity and the structure (form, shape) still remains an unsolved problem.

Ascorbate Oxidase↗

Light-induced resetting of a circadian clock is mediated by a rapid increase in frequency transcript.

To understand how light entrains circadian clocks, we examined the effects of light on a gene known to encode a state variable of a circadian oscillator, the frequency (frq) gene. frq is rapidly induced by short pulses of visible light; clock resetting is correlated with frq induction and is blocked by drugs that block the synthesis of protein or translatable RNA. The speed and magnitude of frq induction suggest that this may be the initial clock-specific event in light resetting. Light induction overcomes frq negative autoregulation so that frq expression can remain high in constant light. These data explain how a simple unidirectional signal (light and the induction of frq) may be turned into a bidirectional clock response (time of day-specific advances and delays). This light entrainment model is easily generalized and may be the common mechanism by which the intracellular feedback cycles that comprise circadian clocks are brought into synchrony with external cycles in the real world.

Analysis of Variance↗

Analysis of Arabidopsis PsbQA gene expression in transgenic tobacco reveals differential role of its promoter and transcribed region in organ-specific and light-mediated regulation.

Arabidopsis PsbQ, encoding a 16 kDa protein of the oxygen-evolving complex, is regulated by light and is expressed preferentially in leaf tissues. To analyze the components required for light-regulated and organ-specific expression of PsbQA, several promoter constructs were generated and expressed in tobacco. The 2.2 kb promoter could confer organ-specific expression of the reporter gene, whereas regulatory elements for light-dependent induction could not be located within this promoter and the transcribed region extending up to a second exon, represented by a genomic fragment encompassing the gene. The genomic fragment representing the transcribed region, however, could confer light regulation even on a constitutive promoter, as observed by steady-state mRNA analysis in T0 and T1 tobacco plants. The results obtained have led to the conclusion that regulatory elements for organ-specificity mainly reside in the promoter region whereas the transcribed region of the gene has an important role in light regulation.

5' Untranslated Regions↗

Induction of plasminogen activator by UV light in normal and xeroderma pigmentosum fibroblasts.

Normal and DNA repair-deficient human fibroblasts have been used to study induction of plasminogen activator (PA) by DNA damage. UV light induced the synthesis of PA in skin fibroblasts of all types of xeroderma pigmentosum (XP) in XP heterozygotes and in human amniotic cells. Enzyme induction was, however, not observed in fibroblasts of normal adults. In classical XP, which are deficient in excision repair, PA synthesis occurred in a narrow range of low-UV fluences. In such strains, the level of enzyme produced was correlated with the extent of repair deficiency. UV fluences required for PA induction in XP variants and XP heterozygotes were at least 10 times those inducing enzyme synthesis in excision-deficient XP. Maximum enzyme induction occurred 48 hr after irradiation, and the highest levels of enzyme produced were 15-20 times those of PA baseline levels. Electrophoretic analysis showed that UV irradiation enhances the synthesis of the Mr 60,000 human urokinase-type PA, which is present in low amounts in untreated cells. Our results suggest that PA induction in human cells is caused by unrepaired DNA damage and represents a eukaryotic SOS-like function. In addition, PA induction may provide a sensitive assay for detection of cellular DNA repair deficiencies and identification of XP heterozygotes.

Amniotic Fluid↗

Phytochrome destruction: an apparent requirement for protein synthesis in the induction of the destruction mechanism.

Examination of the phytochrome destruction reaction as a function of age in etiolated oat (Avena sativa L. cv. Garry) seedlings demonstrates that following illumination of 3-day-old shoots there is a lag, not observed in 4- or 5-day-old oats, prior to the onset of destruction. This light-mediated induction of the phytochrome destruction mechanism in 3-day-old shoots is inhibited by chloramphenicol, actinomycin D, and puromycin suggesting that protein synthesis is required. In 4-day-old shoots, actinomycin D and puromycin do not alter the kinetics of destruction while chloramphenicol partially inhibits the process. Thus, the inhibitors have a specific effect on the induction of the destruction mechanism but not its subsequent operation.

Journal Article↗

Role of Protein Synthesis in Regulation of Phycobiliprotein mRNA Abundance by Light Quality in Fremyella diplosiphon.

If green light-acclimated Fremyella diplosiphon cultures are transferred to red light, the transcription from the inducible phycocyanin gene set increases at least 30-fold within 60 minutes. This effect is inhibited completely by the protein synthesis inhibitors chloramphenicol and spectinomycin. Application of chloramphenicol 30 minutes after transfer of cultures to inductive red light prevents further phycocyanin mRNA accumulation within 10 minutes. If red light-acclimated cells are transferred to green light, the phycocyanin transcript level declines by about 70% within 1 hour. Most of the green light-dependent decline results from the rapid cessation of transcription from the PC gene set. Chloramphenicol slows the decline to some extent by decreasing the rate of mRNA degradation in a light-independent manner. The accumulation of phycoerythrin mRNA after transfer of red light-acclimated cells to green light is also inhibited by chloramphenicol. However, there is no red light-dependent mechanism that rapidly halts phycoerythin mRNA synthesis after transfer of cultures from green to red light. Therefore, at least three light-dependent processes are involved in regulating phycobiliproteingene expression: chloramphenicol-sensitive processes required for the activation of phycocyanin and phycoerythrin gene sets and a chloramphenicol-insensitive process which blocks phycocyanin mRNA synthesis after transfer of cells from red to green light.

Journal Article↗

Effect of the absence of heat shock protein 70.1 (hsp70.1) on retinal photic injury.

This study aimed to evaluate the protective effect of heat shock protein70 (hsp70) on retinal photic injuries, and to determine the relationship between hsp70s from hsp70.1 and 70.3. C57BL/6 wild type (hsp70.1 +/+) and knockout type (hsp70.1 -/-) mice from the same littermates were placed in light of 11000 lux for 6 hours, and were sacrificed at 1, 4, 7, and 14 days after stress. H & E staining, immunohistochemistry, and western blot analysis were performed. The hsp70.1 -/- mice exhibited more disarranged and more diffusely destroyed photoreceptors than the hsp70.1 +/+ mice. Hsp70 induction by light in both the hsp70.1 +/+ and hsp70.1 -/- mice peaked at 1 day after light stress. The Hsp70 level in the hsp70.1 +/+ mice reduced slowly and was almost constant for 7 days. However, in the hsp70.1 -/- mice, it decreased rapidly and returned, after 7 days, to a similar level to that prior to light exposure. According to which gene they originate from, hsp70s may play specific roles in protecting the retina against stresses. Hsp70 from the hsp70.1 gene may act as a sustained responder to retinal photic injury.

Animals↗

Heat shock and light activation of a Chlamydomonas HSP70 gene are mediated by independent regulatory pathways.

Induction of HSP70 heat shock genes by light has been demonstrated in Chlamydomonas. Our aim was to establish whether this induction by light is mediated by the heat stress sensing pathway or by an independent signal chain. Inhibitors of cytoplasmic protein synthesis revealed an initial difference. Cycloheximide and other inhibitors of protein synthesis prevented HSP70A induction upon illumination but not during heat stress. Analysis of HSP70A induction in cells that had differentiated into gametes revealed a second difference. While heat shock resulted in elevated HSP70A mRNA levels, light was no longer able to serve as an inducer in gametes. To identify the regulatory sequences that mediate the response of the HSP70A gene to either heat stress or light we introduced a series of progressive 5' truncations into its promoter sequence. Analyses of the levels of mRNA transcribed from these deletion constructs showed that in most of them the responses to heat shock and light were similar, suggesting that light induction is mediated by a light-activated heat shock factor. However, we show that the HSP70A promoter also contains cis-acting sequences involved in light induction that do not participate in induction by heat stress. Together, these results provide evidence for a regulation of HSP70A gene expression by light through a heat shock-independent signal pathway.

Animals↗

Ultraviolet light inactivation of zinc-mediated metallothionein induction in normal and repair-deficient human cells.

Synthesis of the low molecular weight, thiol-rich, metal-binding metallothioneins (MTS) is undetectable in normal human (NF) or xeroderma pigmentosum (XP) fibroblasts grown in the absence of excess ZnCl2. Addition of 200 microM ZnCl2 to the growth medium produces an increased MT synthesis rising from the basal rate to a rate at least 50-fold greater than basal rate within 7 h. MT induction kinetics in confluent and in exponentially growing subconfluent monolayers were indistinguishable. Zn2+-mediated MT induction is sensitive to actinomycin D suggesting that the induction process is under transcriptional control. Ultraviolet light irradiation causes a dose-dependent inactivation of Zn2+-mediated MT induction in both NF and XP cells. Post-irradiation incubation of UV-irradiated cells using liquid holding techniques leads to reactivation of Zn2+-mediated MT induction in NF cells but not in XP cells. These findings suggest the utility of MT induction produce transcription-terminating lesions, and (b) in evaluating cellular repair capacity for this class of DNA lesions.

Cell Line↗

Changes in hydrophilic antioxidant activity in Avena sativa and Triticum aestivum leaves of different age during de-etiolation and high-light treatment.

The steady-state of reactive oxygen species (ROS) in plant cells is controlled by ROS-producing and scavenging agents. A large cellular pool of antioxidant metabolites is involved in their control. Variations in this antioxidant pool may be monitored by measuring changes in hydrophilic antioxidant activity (free radical-quenching activity of water-soluble components) and ascorbic acid levels. The de-etiolation process and induction of light stress in Avena sativa and Triticum aestivum leaves were used as physiological models to study the antioxidant status at different ages. The data showed that five-day-old green plants and de-etiolated plants of the same age have similar hydrophilic antioxidant activity ( approximately 8 mumol ASC equivalents g FW(-1)), which increases during the de-etiolation process. In oat and wheat, young leaves (five days old) had higher antioxidant status (hydrophilic antioxidant activity and ascorbic acid level) than old leaves (10 and 20 days old). High-light treatment caused a decrease in antioxidant status, especially in young leaves. Hydrophilic antioxidant activity and ascorbic acid levels recovered totally or partially after 30 or 60 min in the dark. This capacity also depends on age and species. The ascorbic acid/hydrophilic antioxidant activity ratio is presented as an indicator of antioxidant variations in response to stress, but taking into account the absolute levels of antioxidants.

Antioxidants↗

Dual role of TOC1 in the control of circadian and photomorphogenic responses in Arabidopsis.

To examine the role of the TOC1 (TIMING OF CAB EXPRESSION1) gene in the Arabidopsis circadian system, we generated a series of transgenic plants expressing a gradation in TOC1 levels. Silencing of the TOC1 gene causes arrhythmia in constant darkness and in various intensities of red light, whereas in blue light, the clock runs faster in silenced plants than in wild-type plants. Increments in TOC1 gene dosage delayed the pace of the clock, whereas TOC1 overexpression abolished rhythmicity in all light conditions tested. Our results show that TOC1 RNA interference and toc1-2 mutant plants displayed an important reduction in sensitivity to red and far-red light in the control of hypocotyl elongation, whereas increments in TOC1 gene dosage clearly enhanced light sensitivity. Furthermore, the red light-mediated induction of CCA1/LHY expression was decreased in TOC1 RNA interference and toc1-2 mutant plants, indicating a role for TOC1 in the phytochrome regulation of circadian gene expression. We conclude that TOC1 is an important component of the circadian clock in Arabidopsis with a crucial function in the integration of light signals to control circadian and morphogenic responses.

Arabidopsis↗

Long-term effects of constant light or darkness on chicken pineal hydroxyindole-O-methyltransferase expression: biochemical and cellular aspects.

1. Chickens kept in constant light, as opposed to constant darkness, display a twofold increase in the activity of pineal hydroxyindole-O-methyltransferase (HIOMT), the last acting enzyme in the melatonin pathway. 2. Using an immunological approach, we presently show that this regulation of HIOMT activity reflects changes in the concentration of a single molecular form of the enzyme protein (a 38 kDa polypeptide). Immunohistofluorescence indicates that these concentration changes concurrently affect modified photoreceptors and pinealocyte-like cells in the chicken pineal organ. 3. Together, the present data support the hypothesis that environmental lighting might regulate the expression of the HIOMT gene.

Acetylserotonin O-Methyltransferase↗