Mutagenic DNA repair in Escherichia coli. XVI. Mutagenesis by ultraviolet light plus delayed photoreversal in recA strains

Mutagenesis was demonstrable after delayed photoreversal of UV-irradiated strains carrying a recA deletion indicating that RecA protein is not essential for the misincorporation process that is revealed by delayed photoreversal. Moreover, the data suggest that RecA protein actually depresses misincorporation to varying extents depending on the recA allele. No delayed photoreversal was demonstrable in reA1 or recA56 bacteria unless the lexA102(ind-) allele was also present. It is suggested that the level of these RecA proteins may be lower in the lexA102(ind-) strains thus minimising their depressive effect. Delayed photoreversal mutagenesis in strains carrying the recA441 allele was not affected by either adenine or guanosine plus cytidine, substances which affect the proteolytic activity of RecA441 protein.     

DNA sequence changes in mutations induced by ultraviolet light in the gpt gene on the chromosome of Escherichia coli uvr+ and urvA cells

Summary Sequence changes in mutations induced by ultraviolet light are reported for the chromosomal Escherichia coli gpt gene in almost isogenic E. coli uvr ⁺ and excision-deficient uvrA cells. Differences between the mutagenic spectra are ascribed to preferential removal of photoproducts in the transcribed strand by excision repair in uvr ⁺ cells. This conclusion is confirmed by analysis of published results for genes in both uvr ⁺ and uvr ^– cells, showing a similar selective removal of mutagenic products from the transcribed strand of the E. coli lacI gene and of the lambda phage cl repressor gene. Comparison of these data with published results for ultraviolet mutagenesis of gpt on a chromosome in Chinese hamster ovary cells showed that a mutagenic hot spot in mammalian cells is not present in E. coli; the possibility is suggested that the hot spot might arise from localized lack of excision repair. Otherwise, mutagenesis in hamster cells appeared similar to that in E. coli uvr ⁺ cells, except there appears to be a smaller fraction of single-base additions and deletions (frameshifts) in mammalian than in bacterial cells. Phenotypes of 6-thioguanine-resistant E. coli showed there is a gene (or genes) other than gpt involved in the utilization of thioguanine by bacteria.     

A proteomic analysis of Helicoverpa armigera adults after exposure to UV light irradiation

Ultraviolet (UV) light (blacklight), which emits UV in the range of 320-400 nm, has been used worldwide in light trapping of insect pests. To gain a better understanding of the response of Helicoverpa armigera adults to UV light irradiation, we carried out a comparative proteomic analysis. Three-day-old adults were exposed to UV light for 1 h. Total proteins were extracted and separated by two-dimensional gel electrophoresis. More than 1200 protein spots were reproducibly detected, including 12 that were more abundant and 21 less abundant. Mass spectrometry analysis and database searching helped us to identify 29 differentially abundant proteins. The identified proteins were categorized into several functional groups including signal transduction, RNA processing, protein processing, stress response, metabolisms, and cytoskeleton structure, etc. This study is the first analysis of differentially expressed proteins in phototactic insects under UV light irradiation conditions and gives new insights into the adaptation mechanisms responsive to UV light irradiation stress.     

Multiplicity reactivation of reovirus particles after exposure to ultraviolet light

McClain, Mary E. (California State Department of Public Health, Berkeley), and Rex S. Spendlove. Multiplicity reactivation of reovirus particles after exposure to ultraviolet light. J. Bacteriol. 92:1422-1429. 1966.-Exposure of reovirus suspensions to moderate doses of ultraviolet light results in essentially exponential inactivation of infectivity to survivals of 10(-2) to 10(-3). With suspensions of sufficiently high particle concentration, larger doses of ultraviolet light (6 to 12 min) are associated with multiplicity reactivation (MR) which is demonstrable both by immunofluorescent-cell count and by plaque assay in FL human amnion cells. Similar effects are produced by photodynamic inactivation in the presence of proflavine, but not by thermal inactivation at 50 C. All three reovirus types exhibit MR under appropriate conditions, and all three interact in mixed ultraviolet suspensions with high efficiency. Progeny from FL cells infected under conditions of MR were as infectious as those of unirradiated inocula, with yields per cell ranging from 10(4) to 4 x 10(4) infective units.     

Using ultraviolet light for improved antifouling performance on ship hull coatings

Abstract

A two-part study was designed to investigate the efficacy of using UVC to prevent biofouling in the context of ship hull coatings. The first study determined the frequency of UVC required for a coating that does not have any additives (epoxy). It was found that 1?min/day was effective at preventing hard fouling but not biofilm development. The second study addressed several variables: coating type (epoxy, copper, fouling release), frequency of UVC (no exposure, continuous exposure, 1min/6h, 1?min/day), and distance from the lamp (25 and 50?mm). Continuous UVC exposure resulted in no biofouling settlement but it did damage the copper coating. Intermittent UVC exposure was effective at preventing biofouling recruitment to both the copper and the fouling release coatings. Variations were observed with regards to the fouling composition, especially biofilms, sedimentary tubeworms and barnacles, suggesting tolerances within the community.     

Study on the synergetic degradation of chitosan with ultraviolet light and hydrogen peroxide

Chitosan was effectively degraded by hydrogen peroxide under irradiation with ultraviolet light. The existence of a synergetic effect on the degradation was demonstrated by means of viscometry. In addition, the optimal conditions of degradation were determined on the basis of orthogonal tests. The structure of the degraded product was characterized by Fourier-transform infrared spectra (FTIR) analysis and diffuse reflectance spectra (DRS) analysis. The mechanism of the degradation of chitosan was correlated with cleavage of the glycosidic bond.     

Effects of ultraviolet radiation and sunlight on the entomogenous nematode,Neoaplectana carpocapsae

Infective-stage juveniles of Neoaplectana carpocapsae were acutely sensitive to short uv radiation (254 nm) and natural sunlight. High nematode mortality, although delayed, accompanied uv exposure. Irradiation rapidly reduced nematode pathogenicity, so that nematodes exposed for 7 min were unable to cause lethal infections in Galleria mellonelia larvae. Moreover, the median survival time of Galleria larvae increased progressively as nematode exposure to uv was lengthened. Inhibition of nematode reproduction and development was noted at exposure periods longer than 2.45 and 5 min, respectively. However, irradiation did not appear to affect juvenile motility. Exposure to direct sunlight also reduced pathogenicity, in a range from 6.9 to 94.9% at 30 and 60 min of exposure, respectively. Long uv (366 nm) did not affect juveniles at the exposures tested.     

Childhood exposure to ultraviolet radiation and harmful skin effects: epidemiological evidence

We review the general amount and patterns of exposure to solar ultraviolet (UV) radiation that children and teenagers experience and the spectrum of UV-related skin damage that can occur as a result. Data about the amount of solar UV received by children and teenagers are relatively few but suggest that around 40–50% of total UV to age 60 occurs before age 20. Among white children, those with the palest complexions suffer the most damage. Comparisons of prevalence and incidence of outcomes in children and teenagers sharing common ancestry, but living at different latitudes, show that prevalence rates of photoaging and melanocytic naevi are higher in Australian compared with British children, and similarly for melanoma. Genetic risk for the majority of the melanomas in teens is a function of genes controlling naevus propensity and pigmentation in the skin. High numbers of naevi and freckles, red hair, blue eyes, inability to tan, as well as a family history are the primary determinants of melanoma among adolescents. Beyond the signs of skin damage seen in children are the latent effects observed later in adulthood. Childhood is believed to be a susceptible window for long-term harmful effects of UV, as evidenced by clear differences in skin cancer risk between child and adult migrants from high to low latitudes. Effective UV radiation protection from childhood is necessary to control both immediate and long-term harmful effects on children’s skin.     

Inhibition and recovery of DNA synthesis in human cells after exposure to ultraviolet light

The inhibition of DNA synthesis in normal human cells by UV is a complex function of fluence because it has several causes. At low fluences, inhibition of replicon initiation is most important. This is made clear by the fact that it occurs to a lesser degree in cells from patients with ataxia telangiectasia (AT). Assuming that only leading strand synthesis is blocked by UV-induced lesions, single lesions between replicons in parental strands for leading strand synthesis inhibit DNA synthesis by acting as temporary blocks until they are replicated by extension of the lagging strand of the adjacent replicon. A more severe inhibition occurs when two lesions are induced between adjacent growing replicons, because one in four possible configurations may result in a long-lived unreplicated region (LLUR). In the absence of excision repair, these may eventually be replicated by activation of an otherwise unused origin within the LLUR. The frequency of LLURs increases steeply with fluence. Activation of normally unused origins to replicate LLURs may facilitate recovery from inhibition of DNA synthesis, but repair of lesions is probably more important. In excision-repair-defective cells, an LLUR without an origin to initiate its replication may be a lethal lesion.     

Acidification induces Bax translocation to the mitochondria and promotes ultraviolet light-induced apoptosis

It has been suggested that Bax translocation to the mitochondria is related to apoptosis, and that cytosol acidification contributes to apoptosis events. However, the mechanisms remain obscure. We investigated the effect of acidification on Bax translocation and on ultraviolet (UV) light-induced apoptosis. The Bax translocation assay in vitro showed that Bax translocated to the mitochondria at pH 6.5, whereas no Bax translocation was observed at pH 7.4. VHDBB cells expressing the GFP-Bax fusion protein were treated for 12 h with a pH 6.5 DMEM medium, nigericin (5 μg/ml) and UV light (50 J/cm²), separately or in combination, and Bax translocation to the mitochondria was determined by SDS-PAGE and Western blot, and apoptotic cell death was detected by flow cytometry. The results showed that some of the Bax translocated to the mitochondria in the cells treated with the normal medium, nigericin and UV in combination, whereas all of the Bax translocated to the mitochondria in the cells treated with the pH 6.5 medium, nigericin and UV in combination. In VHDBB cells treated for 12 h with nigericin, UV alone, and UV and nigericin in combination, the respective rates of apoptotic cell death were 25.08%, 33.25% and 52.88%. In cells treated with pH 6.5 medium and nigericin, pH 6.5 medium and UV, and pH 6.5 medium, nigericin and UV in combination, the respective rates of apoptotic cell death increased to 37.19%, 41.42% and 89.44%. Our results indicated that acidification induces Bax translocation from the cytosol to the mitochondria, and promotes UV lightmediated apoptosis. This suggests that there is a possibility of improving cancer treatment by combining acidification with irradiation or chemotherapeutic drugs.     

Induction of phr gene expression by irradiation of ultraviolet light in Escherichia coli

Summary To measure the degree of phr gene induction by DNA-damaging agents, the promoter region was fused to the coding region of the lacZ gene in plasmid pMC1403. The new plasmids were introduced into Escherichia coli cells having different repair capabilities. More efficient induction of phr gene expression was detected in a uvrA ^– strain as compared with the wild-type strain. In addition, obvious induction was detected in uvrA ^– cells treated by 4-nitroquinoline 1-oxide and mitomycin C. Nalidixic acid, an inhibitor of DNA gyrase, also induced phr gene expression. In contrast, little induced gene expression was noted in UV-irradiated lexA ^– and recA ^– strains. It is suggested from these results that induction of the phr gene is one of the SOS responses. Possible nucleotide sequences which could be considered to constitute an SOS box were found at the regulator region of the phr gene.Abbreviations phr photoreactivation - UV ultraviolet light - 4NQO 4-nitroquinoline 1-oxide - MMC mitomycin C - PRE photoreactivating enzyme - E. coli Escherichia coli     

Terbium fluorescence characteristics of cultured neurons using ultraviolet microspectrofluorometry

The fluorescence emission intensity of terbium is enhanced upon the binding of Tb³⁺ to cultured mouse spinal cord and dorsal root ganglion neurons, via nonradiative resonant energy transfer from membrane proteins. The relative fluorescence intensities of Tb³⁺ bound to dorsal root ganglion neurons were considerably greater than that of Tb³⁺ bound to large multipolar spinal cord neurons. The cell bodies of the dorsal root ganglion neurons were completely covered in a dense fluorescent blanket, whereas the fluorescence from the spinal cord soma presented a discontinuous pattern. The neurites of the spinal cord neuron were speckled with bright patches of Tb³⁺ fluorescence. A high concentration of Ca²⁺ reduced the relative fluorescence intensity of the Tb³⁺ -neuron complex. It is suggested that Tb³⁺ binds to Ca²⁺ -binding sites on the surface membrane of neurons.     

Induction of sister chromatid exchanges in xeroderma pigmentosum cells after exposure to ultraviolet light

The role of DNA repair mechanisms in the induction of sister chromatid exchanges (SCE) after exposure to ultraviolet radiation was investigated in xeroderma pigmentosum cells. Cells from different excision-deficient XP strains, representing the 5 complementation groups in XP, A, B, C, D and E, and from excision-proficient XP variant strains were irradiated with low doses of UVR (0-3.5 J/m2). The number of SCE was counted after two cycles in the presence of BUdR. In cells of the complementation groups A, B, C and D the number of SCE was significantly higher than in UV-exposed control cells. The frequencies of SCE in group E cells and in XP varient cells were not different from those in control cells. Treatment with caffeine (0-200 microgram/ml) did not result in a different response of variant cells compared with normal cells. A simple correlation between SCE frequency and residual excision-repair activity was not observed. The response of the excision-repair deficient cells suggest that unrepaired damage, produced by UVR is involved in the production of SCE.     

Mutagenesis and mutation transfer induced by ultraviolet light in plasmid-cloned DNA

We describe here simple techniques for increasing the frequency of UV-induced mutations in a DNA fragment cloned in plasmid pBR322. Irradiation of both the host and the plasmid DNA before transformation is necessary to produce new mutations in the plasmid DNA, presumably because the UV-damaged pBR322 replicon cannot efficiently induce the error-prone repair pathway of Escherichia coli. In contrast, U V irradiation of the plasmid DNA alone before transformation primarily causes the transfer of preexisting mutations from the host chromosome to homologous DNA present in the plasmid. The only other kind of mutants obtained were large deletions of the plasmid DNA. Two chromosomal mutations from the host galK gene and one from the lacZ gene have been transferred to the plasmid by UV irradiation of the plasmid DNA alone. The technique can thus be of general use.