Induction of mouse epidermal ornithine decarboxylase activity and DNA synthesis by ultraviolet light.
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Regulation of glucoamylase production in photoinduced A. niger under blue light were investigated. The results showed that continuous illumination of blue light was effective in promotion of conidiation, leading to more glucoamylase production due to the need of sporulation in A. niger. It was clear that 36h-old mycelium grown in dark was in the developmental stage of formation of conidiophore stalk and was most sensitive to BL A. niger cultures exhibited some competence stage to respond to blue light over a period of 36 h when early conidiophore stalk formed in darkness, and photoinduction at this critical period resulted in relatively higher yield of glucoamylase. The fluence-response data generated with blue light indicated that the optimum fluence required for the photomorphogenetic response in A. niger was no less than 450 (mol/m2 x s) to saturate the photoinduction system. The amount of glucoamylase gene (G1) transcripts accumulated during illumination with blue light was more than that in total darkness, whereas the threshold for G1 induction during illumination with increased gradient light was higher than that exposed directly to light of constant fluence, indicating the existence of a light adaptation mechanism for glucoamyalse production in response to blue light. Nevertheless, both high and low light were able to promote glucoamylase producing. Furthermore, suppression subtractive hybridization experiment revealed that some respiratory chain redox enzymes in mitochondria including alternative oxidase as well as sulfhydryl oxidase participated the photoresponse in A. niger and consequently influenced the metabolism. The results support a possibility of designing strategies to improve glucoamylase yield by application of blue light.
The tumor necrosis factor family ligands, LIGHT (lymphotoxin like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes), 4-1BBL and CD70, are found in the same gene cluster on mouse chromosome 17. Although the roles of 4-1BB-4-1BBL and CD27-CD70 interactions in anti-viral T cell responses have been well established, the role of LIGHT in T cell activation/expansion in vivo is less clear. Under conditions that were previously employed to demonstrate a role for 4-1BBL in CD8+ T cell memory, wild-type and LIGHT-/- mice were infected with influenza A virus and primary and memory/recall responses were measured at various time points thereafter. Neither primary expansion nor memory/recall CD8+ T cell responses were affected by the absence of LIGHT, as measured up to 2 months post-infection. CD4+ T cell responses were also unaffected by LIGHT deficiency. Furthermore, we found that LIGHT played no role in the induction of influenza-specific IgG1 and IgG2a serum antibodies. Taken together, these data suggest that LIGHT is dispensable for the acquired immune response to influenza virus in mice with no effect on the induction, maintenance or reactivation of CD8+ T cell memory.
The cumacean Dimorphostylis asiatica (Crustacea) exhibits a circatidal swimming activity rhythm. The animals were exposed to a 12.5 hr sinusoidal change of hydrostatic pressure of 0.3 atm amplitude in the laboratory. Under constant dark conditions, most of the specimens were entrained to a daily bimodal swimming activity rhythm by the hydrostatic pressure cycle. A small number of individuals exhibited a unimodal daily rhythm, with no apparent entraining from the administered cycles. A marked feature was a flexible phase relationship between the entrained daily bimodal rhythm and the hydrostatic pressure cycles: the swimming activity of most of the specimens occurred around the pressure-decreasing phase, but for a small number of individuals it coincided with the pressure-increasing phase. Such flexibility suggests a weak entraining effect of hydrostatic pressure on the circatidal rhythm of this species. When exposed to 24 hr light-dark cycles and a hydrostatic pressure cycle simultaneously, the specimens exhibited a rhythmic activity entrained by the hydrostatic pressure cycle during the dark period, which closely resembles the temporal activity pattern of this species in the field. The light cycles entrained the swimming activity via direct inhibition and induction of activity (i.e., masking). Under light-dark conditions, the specimens exhibited activity on the pressure-increasing phase more frequently compared with specimens kept in constant darkness.
In a common garden, we investigated genetic and environmental influences on alkaloid production using Sanguinaria canadensis as a model. Nutrient and shade regimes were applied to replicated clones over one growing season, and induction of alkaloid production in bloodroot was tested on a whole-plant basis using jasmonic acid as an elicitor. Alkaloid concentrations increased with decreasing light intensity and fertilizer levels. Induction was not achieved by foliar application of jasmonic acid. Genetic influences represented by clone effects may be indicated by variation in alkaloid concentration by clone, but this experimental design did not allow us to distinguish genetic from pre-experiment environmental influences on the rhizomes.
Neonatal treatment of rodents by intraperitoneal injections of monosodium glutamate (MSG) destroys many retinal ganglion cells whose neurons belong to the circadian system; howertheless, adults always synchronize their locomotor activity rhythm (LAR) to the light/dark cycle. Recent studies have shown that light-induced phase shifts of LAR are associated with the c-fos induction in suprachiasmatic nuclei (SCN) of nocturnal rodents. In this study, the circadian system was analyzed in treated and control hamsters maintained in constant darkness and exposed to light at circadian times (CTs) 13 and 18 during subjective night, 1 and 6 h after the onset of LAR. The period of the LAR and delay (CT13) and advance (CT18) phase shifts of LAR were not significantly different between MSG-treated and control hamsters. Temporospatial variations of Fos induction after light exposure were similar in both MSG-treated and control hamsters although the total number of Fos immunoreactive (Fos-ir) nuclei in the SCN was always lower in treated hamsters. However, the decrease in Fos-ir was significant only for the caudal third of the SCN of treated hamsters, the part where retinal afferents are most dense. The effect of light exposure on Fos expression in SCN of MSG-treated and control hamsters was the same at CT13 and CT18: (1) Fos-ir nuclei were significantly more numerous at CT18 than at CT13 in the rostral SCN; (2) dorsal Fos-ir cells were observed in the SCN only at CT18; (3) a ventral subgroup expressed Fos protein in intermediate SCN only at CT13. This study demonstrates that MSG-treatment does not significantly modify the phase-shifting effects of light on either the LAR or the associated cellular activation.
Dark-grown hypocotyls of a starch-deficient mutant (NS458) of tobacco (Nicotiana sylvestris) lack amyloplasts and plastid sedimentation, and have severely reduced gravitropism. However, gravitropism improved dramatically when NS458 seedlings were grown in the light. To determine the extent of this improvement and whether mutant hypocotyls contain sedimented amyloplasts, gravitropic sensitivity (induction time and intermittent stimulation) and plastid size and position in the endodermis were measured in seedlings grown for 8 d in the light. Light-grown NS458 hypocotyls were gravitropic but were less sensitive than the wild type (WT). Starch occupied 10% of the volume of NS458 plastids grown in both the light and the dark, whereas WT plastids were essentially filled with starch in both treatments. Light increased plastid size twice as much in the mutant as in the WT. Plastids in light-grown NS458 were sedimented, presumably because of their larger size and greater total starch content. The induction by light of plastid sedimentation in NS458 provides new evidence for the role of plastid mass and sedimentation in stem gravitropic sensing. Because the mutant is not as sensitive as the WT, NS458 plastids may not have sufficient mass to provide full gravitropic sensitivity.
Prophage S2 could be induced by psoralen plus near-UV light (PNUV) from a wild-type strain of Haemophilus influenzae, from UV light-sensitive strains uvr-1 and uvr-2 and PNUV-sensitive strains PSO1 amd PSO7, but not from a recombination-deficient strain, rec-1. The levels of prophage induction were comparable in the wild type and an ATP-dependent DNase-deficient strain, KW31, even though the PNUV-induced degradation in the latter strain was considerably lower. Prophage induction could be observed even with chloramphenicol present before, during, and 30 min after PNUV treatment.
BACKGROUND/AIMS: Galectin-3 is an interesting intracellular lectin that appears to be involved in numerous physiological processes. We have analyzed expression of galectin-3 in glioblastoma cells exposed to heat-shock, alkylating agents, UV-C radiation and subculturing (trypsinization). METHODS: Protein levels of galectin-3 were measured by western-blot analysis using M3/38 monoclonal antibody. The involvement of transcription factors NF-kappaB and Jun in the induction of galectin-3 was addressed using specific inhibitor of NF-kappaB (zL(3)-vs) and antisense-jun oligonucleotides. RESULTS: Exposure of cells to heat-shock or subculturing (trypsinization) decreased levels of galectin-3 to approximately 50%. Alkylating damage and UV-C irradiation caused an increase in the expression of galectin-3. Both inhibition of Jun by antisense-jun oligonucleotides, and inhibition of NF-kappaB by specific proteasomal inhibitor attenuated the induction of galectin-3 by UV-light, but with somewhat different kinetics. CONCLUSIONS: We have found that different forms of cellular stress have different effects on the expression of galectin-3. Heat-shock and subculturing decrease, while alkylating agents and UV-light increase galectin 3. NF-kappaB and Jun were shown to be involved in the induction of galectin-3 by UV-light, which is a first demonstration that these transcriptional factors are involved in the regulation of galectin-3 expression.
Nuclear genes encoding plastid ribosomal proteins are more highly expressed in leaves than in roots. This leaf-specific induction seems to be light-independent. We have previously characterized a spinach nuclear factor S1F binding to a cis-element within the rps1 promoter, which negatively regulates both the rps1 and the cauliflower mosaic virus 35S promoters in transient expression assays. Here, we show that the S1F binding site is related to but different from the light-responsive Box II of the pea rbcS-3A promoter, which is recognized by the nuclear factor GT-1. Transgenic plant analyses showed that the S1F site tissue-specifically represses the rps1 promoter in roots as well as in etiolated seedlings. We suggest that the GT-1-related S1F binding site is responsible, at least in part, for the transcriptional repression of rps1 in nonphotosynthetic tissues such as roots.
The gene con-10 of Neurospora crassa is expressed preferentially during conidiation and following illumination of vegetative mycelia with blue light. In this study we have examined the segmental locations of the genetic elements associated with con-10 that are responsible for light and developmental expression. A translational fusion was prepared between the initial segment of con-10 and Escherichia coli lacZ. Deletions were then introduced into the con-10 upstream region associated with this translational fusion. Each construct was integrated at the his-3 locus of N. crassa by transformation and homologous recombination. Photoinduction of mycelia containing the translational fusion with the intact upstream region revealed a two phase stimulus-response curve. Exposure to light for as little as 5 sec induced a transcriptional response. Following this initial induction, a period of 15 min in the dark or light was required for appearance of a second phase response. Only a brief light treatment was necessary for induction of the second phase response. Deletions within the upstream region altered normal light and developmental expression of constructs containing the con-10-lacZ translational fusion. The deleted segments appear to contain a mycelial repression site, two conidiation activation sites, and two dark repression sites. A repeated 17-bp sequence acted as a transcriptional enhancer. One copy of this enhancer is in the upstream region. The second copy, with the opposite orientation, is located in the first con-10 intron. The enhancer was required for proper mycelial and conidial expression of the con-10-lacZ fusion. The initial 10 bp of this enhancer sequence were sufficient to restore conidial expression to a deletion construct lacking both copies of the 17-bp repeat. Proteins were detected in extracts of mycelia and conidia that specifically bound to the enhancer sequence in vitro. Our findings suggest that conidiation-specific and mycelial-specific expression of con-10 requires the action of several factors acting independently and/or in concert at distinct sites located in the regulatory regions for con-10.
Salmonella typhosa hybrids able to adsorb lambda were obtained by mating S. typhosa recipients with Escherichia coli K-12 donors. After adsorption of wild-type lambda to these S. typhosa hybrids, no plaques or infective centers could be detected. E. coli K-12 gal(+) genes carried by the defective phage lambdadg were transduced to S. typhosa hybrids with HFT lysates derived from E. coli heterogenotes. The lysogenic state which resulted in the S. typhosa hybrids after gal(+) transduction differed from that of E. coli. Ability to produce lambda, initially present, was permanently segregated by transductants of the S. typhosa hybrid. S. typhosa lysogens did not lyse upon treatment for phage induction with mitomycin C, ultraviolet light, or heat in the case of thermoinducible lambda. A further difference in the behavior of lambda in Salmonella hybrids was the absence of zygotic induction of the prophage when transferred from E. coli K-12 donors to S. typhosa. A new lambda mutant class, capable of forming plaques on S. typhosa hybrids refractory to wild-type lambda, was isolated at low frequency by plating lambda on S. typhosa hybrid WR4254. Such mutants have been designated as lambdasx, and a mutant allele of lambdasx was located between the P and Q genes of the lambda chromosome. Plaques were formed also on the S. typhosa hybrid host with a series of lambda(i21) hybrid phages which contain the N gene of phage 21. The significance of these results in terms of Salmonella species as hosts for lambda is discussed.
We developed a transcript profiling methodology to elucidate expression patterns of the cyanobacterium Synechocystis sp. strain PCC 6803 and used the technology to investigate changes in gene expression caused by irradiation with either intermediate-wavelength UV light (UV-B) or high-intensity white light. Several families of transcripts were altered by UV-B treatment, including mRNAs specifying proteins involved in light harvesting, photosynthesis, photoprotection, and the heat shock response. In addition, UV-B light induced the stringent response in Synechocystis, as indicated by the repression of ribosomal protein transcripts and other mRNAs involved in translation. High-intensity white light- and UV-B-mediated expression profiles overlapped in the down-regulation of photosynthesis genes and induction of heat shock response but differed in several other transcriptional processes including those specifying carbon dioxide uptake and fixation, the stringent response, and the induction profile of the high-light-inducible proteins. These two profile comparisons not only corroborated known physiological changes but also suggested coordinated regulation of many pathways, including synchronized induction of D1 protein recycling and a coupling between decreased phycobilisome biosynthesis and increased phycobilisome degradation. Overall, the gene expression profile analysis generated new insights into the integrated network of genes that adapts rapidly to different wavelengths and intensities of light.
Three methotrexate (MTX)-resistant clones selected at 200 nM MTX have been isolated from UV-irradiated V79 cells and were marked M1, M2 and M3. Two further clones were also isolated by gradually exposing M1 to higher concentrations of MTX and marked M4 and M5, isolated at 1200 nM and 2400 nM MTX respectively. The frequency of the induction of MTX-resistant clones by UV light was found to increase significantly on treatment of the irradiated cells with either 3-aminobenzamide or caffeine. The resistant clones had the same sensitivity as the parental cells to UV light, but were resistant to gamma-rays, MNNG and H2O2. In MTX-resistant cells low-dose H2O2-induced repair function was absent. UV-induced mutation at the HGPRT locus was the same in MTX-resistant as sensitive cells, whereas for MNNG-induced mutation it was higher in resistant cells. The rate of SCE induction by UV light was higher in MTX-resistant cells compared to the parental ones, but was not significantly different for induction by MNNG and H2O2.
Light-induced germination of Anemia spores can be inhibited by AMO-1618, a selective inhibitor of gibberellin biosynthesis. The inhibitor has no effect on gibberellin-induced dark germination and its inhibition of light-induced germination can be reversed by supplying gibberellin. Barley-endosperm bioassay of concentrates of medium in which spores are imbibed in light reveals the presence of substances with gibberellin-like activity; assay of medium from dark-imbibed spores does not. Simultaneous exposure of spores to sub-optimal levels of light and gibberellin leads to additivity of effect on germination level. Uptake of labeled gibberellin by spores in light is similar to that in darkness. The implications of these findings for the light-dependent synthesis of a gibberellin-like germination substance are discussed. The bearing of the observations upon understanding the interaction of light and gibberellins in seeds of higher plants is considered.
The activity of ultraviolet (UV) light to induce unscheduled DNA synthesis (UDS) was investigated in hairless mouse epidermis by means of an in vivo-in vitro assay using a liquid scintillation counting method. Groups of three to five 8-week-old female hairless mice were irradiated with UV-B or UV-A, then skin samples were taken and cultured individually in medium containing [3H]thymidine with or without hydroxyurea (HU) for 2 hr. DNA of the epidermis was extracted, and incorporation of [3H]thymidine and the DNA content were determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS was judged in terms of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. UV-B increased the UDS index 1 hr after irradiation of 500 J/m2, which corresponds to approximately 1 minimal erythema dose or 1 minimal edema dose, and showed a dose-dependent increase up to 17-fold in the UDS index at irradiation doses of 500 to 2,000 J/m2. In a time-course study, UV-B also increased replicative DNA synthesis (RDS) 48 hr after irradiation at 1,000 J/m2. On the other hand, UV-A did not increase the UDS index at irradiation doses of 2 x 10(5) to 8 x 10(5) J/m2. These results show that induction of UDS by UV irradiation depends on wavelength and an increase of RDS in the epidermis exposed to UV-B irradiation appears after induction of UDS.
The chicken pineal gland contains the autonomous circadian oscillator together with the photic-input pathway. We searched for chicken pineal genes that are induced by light in a time-of-day-dependent manner, and isolated chicken homolog of bZIP transcription factor E4bp4 (cE4bp4) showing high similarity to vrille, one of the Drosophila clock genes. cE4bp4 was expressed rhythmically in the pineal gland with a peak at very early (subjective) night under both 12-h light/12-h dark cycle and constant dark conditions, and the phase was nearly opposite to the expression rhythm of cPer2, a chicken pineal clock gene. Luciferase reporter gene assays showed that cE4BP4 represses cPer2 promoter through a E4BP4-recognition sequence present in the 5'-flanking region, indicating that cE4BP4 can down-regulate the chick pineal cPer2 expression. In vivo light-perturbation studies showed that the prolongation of the light period to early subjective night maintained the high level expression of the pineal cE4bp4, and presumably as a consequence delayed the onset of the induction of the pineal cPer2 expression in the next morning. These light-dependent changes in the mRNA levels of the pineal cE4bp4 and cPer2 were followed by a phase-delay of the subsequent cycles of cE4bp4/cPer2 expression, suggesting that cE4BP4 plays an important role in the phase-delaying process as a light-dependent suppressor of cPer2 gene.
To study the mechanism and risk of human skin cancer from solar light, we exposed human skin transplanted to severe combined immunodeficient mice to daily doses of UVB for periods of approximately 2 years. We have succeeded for the first time in inducing cancer and solar (actinic) keratosis in human skin by UVB. Of 18 normal skins exposed to doses of 7.3 x 10(5) to 1.8 x 10(6) J/m2, 14 actinic keratoses (77.8%) and 3 squamous cell carcinomas (16.7%) developed, whereas neither actinic keratosis nor cancer was observed in 15 human skins not exposed to UVB. Each human skin showed a different susceptibility, and skins sensitive for actinic keratosis were also sensitive for cancer induction. Among p53 mutations at various sites, mutation at codon 242 (C TGC --> C CGC; Cys --> Arg) was specifically observed in both skin cancers and actinic keratoses. Furthermore, double or triple mutations were induced in all UVB-induced skin cancers and in three of eight actinic keratoses. Most of the mutations (17 of 20) occurred at dipyrimidine sites.