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Cyclobutane pyrimidine dimers (CPDs) are the main mutagenic DNA photoproducts caused by ultraviolet B (UVB) radiation and represent the major cause of photoaging and skin carcinogenesis. CPD photolyase can efficiently and rapidly repair CPD products. Therefore, they are candidates for the prevention of photodamage. However, these photolyases are not present in placental mammals. In this study, we produced a recombinant photolyase-thymine (rPHO) from Thermus thermophilus (T. thermophilus). The rPHO displayed CPD photorepair activity. It prevented UVB-induced DNA damage by repairing CPD photoproducts to pyrimidine monomers. Furthermore, it inhibited UVB-induced ROS production, lipid peroxidation, inflammatory responses, and apoptosis. UVB-induced wrinkle formation, epidermal hyperplasia, and collagen degradation in mice skin was significantly inhibited when the photolyase was applied topically to the skin. These results demonstrated that rPHO has promising protective effects against UVB-induced photodamage and may contribute to the development of anti-UVB skin photodamage drugs and cosmetic products. [ABSTRACT FROM AUTHOR]

Cyclobutane pyrimidine dimers (CPDs) are the main mutagenic DNA photoproducts caused by ultraviolet B (UVB) radiation and represent the major cause of photoaging and skin carcinogenesis. CPD photolyase can efficiently and rapidly repair CPD products. Therefore, they are candidates for the prevention of photodamage. However, these photolyases are not present in placental mammals. In this study, we produced a recombinant photolyase-thymine (rPHO) from Thermus thermophilus (T. thermophilus). The rPHO displayed CPD photorepair activity. It prevented UVB-induced DNA damage by repairing CPD photoproducts to pyrimidine monomers. Furthermore, it inhibited UVB-induced ROS production, lipid peroxidation, inflammatory responses, and apoptosis. UVB-induced wrinkle formation, epidermal hyperplasia, and collagen degradation in mice skin was significantly inhibited when the photolyase was applied topically to the skin. These results demonstrated that rPHO has promising protective effects against UVB-induced photodamage and may contribute to the development of anti-UVB skin photodamage drugs and cosmetic products.

Marine bacterium Rhodococcus sp. NJ-530 has developed several ultraviolet (UV) adaptive characteristics for survival and growth in extreme Antarctic environment. Rhodococcus sp. NJ-530 DNA photolyase encoded by a 1146 bp photolyase-homologous region (phr) was identified in genome. Quantitative real-time PCR demonstrated that transcriptional levels of phr were highly up-regulated by ultraviolet-B (UV-B) radiation (90 μW·cm−2) and increased to a maximum of 149.17-fold after exposure for 20 min. According to the results of SDS-PAGE and western blot, PHR was effectively induced by isopropyl-β-d-1-thiogalactopyranoside (IPTG) at the genetically engineered BL21(DE3)-pET-32a(+)-phr construct under the condition of 15 °C for 16 h and 37 °C for 4 h. In terms of in vivo activity, compared with a phr-defective E. coli strain, phr-transformed E. coli exhibited higher survival rate under high UV-B intensity of 90 μW·cm−2. Meanwhile, the purified PHR, with blue light, presented obvious photorepair activity toward UV-induced DNA damage in vitro assays. To sum up, studying the mechanisms of Rhodococcus sp. NJ-530 photolyase is of great interest to understand the adaptation of polar bacteria to high UV radiation, and such data present important therapeutic value for further UV-induced human skin and genetic damage diseases. [ABSTRACT FROM AUTHOR]

Photolyases are flavoproteins that repair ultraviolet-induced DNA lesions (cyclobutane pyrimidine dimer or CPD, and pyrimidine (6-4) pyrimidone photoproducts or (6-4)-PPs), using blue light as an energy source. These enzymes are substrate specific, meaning that a specific photolyase repairs either a CPD or a (6-4)-PP. In this work, we produced a class II CPD-photolyase (called as PhrSph98) from the Antarctic bacterium Sphingomonas sp. UV9 by recombinant DNA technology and we purified the enzyme using immobilized metal affinity chromatography. By using an immunochemistry assay, with monoclonal antibodies against CPD and (6-4)-PP, we found that PhrSph98 repairs both DNA lesions. The result was confirmed by immunocytochemistry using immortalized non-tumorigenic human keratinocytes. Results from structure prediction, pocket computation, and molecular docking analyses showed that PhrSph98 has the two expected protein domains (light-harvesting antenna and a catalytic domain), a larger catalytic site as compared with photolyases produced by mesophilic organisms, and that both substrates fit the catalytic domain. The results obtained from predicted homology modeling suggest that the electron transfer pathway may occur following this pathway: Y389-W369-W390-F376-W381/FAD. The evolutionary reconstruction of PhrSph98 suggests that this is a missing link that reflects the transition of (6-4)-PP repair into the CPD repair ability for the class II CPD-photolyases. To the best of our knowledge, this is the first report of a naturally occurring bifunctional, CPD and (6-4)-PP, repairing enzyme. Key points: • We report the first described bifunctional CPD/(6-4)-photoproducts repairing enzyme. The bifunctional enzyme reaches the nuclei of keratinocyte and repairs the UV-induced DNA damage. The enzyme should be a missing link from an evolutionary point of view. The enzyme may have potential uses in the pharmaceutical and cosmetic industries. [ABSTRACT FROM AUTHOR]

Molecular mechanisms underlying insect-pathogenic fungal tolerance to solar ultraviolet (UV) damage have been increasingly understood. This chapter reviews the methodology established to quantify fungal response to solar UV radiation, which consists of UVB and UVA, and characterize a pattern of the solar UV dose (damage) accumulated from sunrise to sunset on sunny summer days. An emphasis is placed on anti-UV mechanisms of fungal insect pathogens in comparison to those well documented in model yeast. Principles are discussed for properly timing the application of a fungal pesticide to improve pest control during summer months. Fungal UV tolerance depends on either nucleotide excision repair (NER) or photorepair of UV-induced DNA lesions to recover UV-impaired cells in the darkness or the light. NER is a slow process independent of light and depends on a large family of anti-UV radiation (RAD) proteins studied intensively in model yeast but rarely in non-yeast fungi. Photorepair is a rapid process that had long been considered to depend on only one or two photolyases in filamentous fungi. However, recent studies have greatly expanded a genetic/molecular basis for photorepair-dependent photoreactivation that serves as a primary anti-UV mechanism in insect-pathogenic fungi, in which photolyase regulators required for photorepair and multiple RAD homologs have higher or much higher photoreactivation activities than do photolyases. The NER activities of those homologs in dark reactivation cannot recover the severe UV damage recovered by their activities in photoreactivation. Future studies are expected to further expand the genetic/molecular basis of photoreactivation and enrich principles for the recovery of insect-pathogenic fungi from solar UV damage., (Copyright © 2024. Published by Elsevier Inc.)

Photolyases are DNA-repairing flavoproteins that are represented in most phylogenetic taxa with the exception of placental mammals. These enzymes reduce the ultraviolet-induced DNA damage; thus, they have features that make them very attractive for dermatological or other medical uses, such as the prevention of human skin cancer and actinic keratosis. In this work, we identified a 50.8 kDa photolyase from the UVC-resistant Antarctic bacterium Hymenobacter sp. UV11. The enzyme was produced by recombinant DNA technology, purified using immobilized metal affinity chromatography and its activity was analyzed using different approaches: detection of cyclobutane pyrimidine dimers (CPDs) by immunochemistry, high-performance liquid chromatography and comet assays using Chinese Hamster Ovary (CHO) and immortalized nontumorigenic human epidermal (HaCat) cells. The information supports that the recombinant protein has the ability to repair the formation of CPDs, on both double- and single-stranded DNA. This CPD-photolyase was fully active on CHO and HaCat cell lines, suggesting that this enzyme could be used for medical or cosmetic purposes. Results also suggest that the UV11 CPD-photolyase uses MTHF as chromophore in the antenna domain. The potential use of this recombinant enzyme in the development of new inventions with pharmaceutical and cosmetic applications is discussed during this work. [ABSTRACT FROM AUTHOR]

This work introduces a new experimental method for the comprehensive description of the physiological responses to light of photosynthetic organisms. It allows the integration in a single experiment of the main established manipulative chlorophyll fluorescence-based protocols. It enables the integrated characterization of the photophysiology of samples regarding photoacclimation state (generating non-sequential light-response curves of effective PSII quantum yield, electron transport rate or non-photochemical quenching), photoprotection capacity (running light stress-recovery experiments, quantifying non-photochemical quenching components) and the operation of photoinactivation and photorepair processes (measuring rate constants of photoinactivation and repair for different light levels and the relative quantum yield of photoinactivation). The new method is based on a previously introduced technique, combining the illumination of a set of replicated samples with spatially separated actinic light beams of different intensity, and the simultaneous measurement of the fluorescence emitted by all samples using an imaging fluorometer. The main novelty described here is the independent manipulation of light intensity and duration of exposure for each sample, and the control of the cumulative light dose applied. The results demonstrate the proof of concept for the method, by comparing the responses of cultures of Chlorella vulgaris acclimated to low and high light regimes, highlighting the mapping of light stress responses over a wide range of light intensity and exposure conditions, and the rapid generation of paired light-response curves of photoinactivation and repair rate constants. This approach represents a chlorophyll fluorescence 'protocol of everything', contributing towards the high throughput characterization of the photophysiology of photosynthetic organisms. [ABSTRACT FROM AUTHOR]

• QMRA model was developed for photorepair after UV disinfection in WASH contexts. • Photorepair kinetics were modelled using reproducible fluence-based determinations. • Competing photorepair and solar disinfection kinetics impact risk. • UV fluences ≥ 16 mJ/cm2 minimize risk for POU and packaged water consumption. • Photorepair risk is low in UV treated packaged water when transport or storage is < 60 min. Ultraviolet disinfection is a promising solution for decentralized drinking water systems such as communal water taps. A potential health risk is enzymatic photorepair of pathogens after UV disinfection, which can result in regrowth of pathogens. Even though photorepair is a known issue, no formal risk assessments have been conducted for photorepair after UV disinfection in drinking water. The main objective was to construct a quantitative microbial risk assessment (QMRA) of photorepair after UV disinfection of drinking water in a decentralized system. UV disinfection and photorepair kinetics for E. coli were modelled using reproducible fluence-based determinations. Impacts of water collection patterns, and wavelength-dependent water container material transmittance, sunlight intensity, and photorepair enzyme absorbance were quantified. After UV disinfection by 16 or 40 mJ/cm2 of < 5-log microorganisms per L, risk of infection did not exceed 1-in-10,000 under conditions permitting E. coli photorepair. Risk from photorepair was less than 1-in-10,000 for photorepair light exposure < 0.75 h throughout the day for UV fluence 16 mJ/cm2 or greater. UV disinfection followed by solar disinfection surpassing photoreactivation during storage reduced risk below 1-in-10,000 for photorepair light exposure > 2.5 h between modelled times of 9 AM – 3 PM. The model can be expanded to other pathogens as UV fluence and photorepair fluence response kinetics become available, and this QMRA can be used to inform the placement of community water access points to reduce risk of photorepair and ensure adequate shelf life of UV disinfected water under safe storage conditions. [Display omitted] [ABSTRACT FROM AUTHOR]

High doses of ultraviolet-B (UV-B; 280-315 nm) radiation can have detrimental effects on plants, and especially damage their DNA. Plants have DNA repair and protection mechanisms to prevent UV-B damage. However, it remains unclear how DNA damage and tolerance mechanisms vary among field species. We studied DNA damage and tolerance mechanisms in 26 species with different functional groups coexisting in two moorlands at two elevations. We collected current-year leaves in July and August, and determined accumulation of cyclobutane pyrimidine dimer (CPD) as UV-B damage and photorepair activity (PRA) and concentrations of UV-absorbing compounds (UACs) and carotenoids (CARs) as UV-B tolerance mechanisms. DNA damage was greater in dicot than in monocot species, and higher in herbaceous than in woody species. Evergreen species accumulated more CPDs than deciduous species. PRA was higher in Poaceae than in species of other families. UACs were significantly higher in woody than in herbaceous species. The CPD level was not explained by the mechanisms across species, but was significantly related to PRA and UACs when we ignored species with low CPD, PRA and UACs, implying the presence of another effective tolerance mechanism. UACs were correlated negatively with PRA and positively with CARs. Our results revealed that UV-induced DNA damage significantly varies among native species, and this variation is related to functional groups. DNA repair, rather than UV-B protection, dominates in UV-B tolerance in the field. Our findings also suggest that UV-B tolerance mechanisms vary among species under evolutionary trade-off and synergism. [ABSTRACT FROM AUTHOR]

Actinic keratosis (AK) or lesions of epidermal dysplasia occurring in skin chronically exposed to solar radiation is very prevalent in lighter skin persons, with chronic long-term sun exposure being the major risk factor. With an aging population it is expected that the prevalence of AK will further increase. AK can progress to nonmelanoma skin cancer (NMSC) and is a public health concern. Six leading dermatologists with expertise in AK and NMSC from Germany met to discuss the nature of the disease and the prevention and treatment strategies available to dermatologists today. While cosmetic sunscreen products form an essential element of sun protection strategies, they are not adequate when damage has already been inflicted. Newly developed products of the medical device category offer DNA repair function paired with high sun protection factor (SPF) UV protection. An adjuvant treatment algorithm for various risk levels of AK was developed. For patients with low and moderate risk, sunscreen only is recommended. For patient groups with high and very high risk, a very high photoprotection and photorepair action (DNA repair enzymes) in medical device products all year round is recommended. © 2015 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Plants require solar energy to grow through oxygenic photosynthesis; however, when light intensity exceeds the optimal range for photosynthesis, it causes abiotic stress and physiological damage in plants. In response to high light stress, plants initiate a series of signal transduction from chloroplasts to whole cells and from locally stressed tissues to the rest of the plant body. These signals trigger a variety of physiological and biochemical reactions intended to mitigate the deleterious effects of high light intensity, such as photodamage and photoinhibition. Light stress protection mechanisms include chloroplastic Reactive oxygen species (ROS) scavenging, chloroplast and stomatal movement, and anthocyanin production. Photosynthetic apparatuses, being the direct targets of photodamage, have also developed various acclimation processes such as thermal energy dissipation through nonphotochemical quenching (NPQ), photorepair of Photosystem II (PSII), and transcriptional regulation of photosynthetic proteins. Fluctuating light is another mild but persistent type of light stress in nature, which unfortunately has been poorly investigated. Current studies, however, suggest that state transitions and cyclic electron transport are the main adaptive mechanisms for mediating fluctuating light stress in plants. Here, we review the current breadth of knowledge regarding physiological and biochemical responses to both high light stress and fluctuating light stress., Competing Interests: Conflict of interest The authors declare that no conflict of interest exists., (Copyright © 2022 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.)

Zooplankton species have evolved several adaptive strategies to minimize damage caused by exposure to solar ultraviolet radiation, but the environmental conditions favoring one strategy or another are not yet fully understood. Here, I quantified the concentration of photoprotective compounds (carotenoids and mycosporine-like amino acids or MAAs) and assessed the photorepair activity (photolyase assay) in populations of the calanoid copepod, Arctodiaptomus jurisowitchi and the cladocerans, Daphnia himalaya and D. longispina, from five high altitude lakes located in the Himalayan Region (Khumbu Valley, Nepal) between 4890 and 5440 m above sea level. The concentration and diversity of MAAs were low in copepods, as well as in seston samples. Significant differences in the concentration of MAAs among the five copepod populations were largely explained (96%) by the lake depth refuge (i.e., the fraction of the water column to which 1% of the surface UVR at 320 nm penetrates). Concentrations of carotenoids (mostly free astaxanthin) in copepods were among the highest reported in the literature. Similar to MAAs, the carotenoid concentration was inversely related to the lake depth refuge. The lowest concentration of photoprotective compounds in copepods was observed in a turbid glacier lake, whereas the highest was found in a shallow water body dominated by a benthic mat of filamentous green algae. Except for the presence of melanin in D. himalaya, no other photoprotective compounds were found in cladocerans. The assay of photolyase activity in A. jurisowitchi and D. himalaya suggested the absence of a photorepair mechanism. The results of this study indicate that the copepod populations from this relatively pristine alpine region rely mainly on the accumulation of carotenoids to minimize damage by UV radiation, a pattern that strongly contrasts with what is known for copepods from other alpine lakes, for instance, in the Alps. I hypothesize that this difference is attributed to nitrogen limitation of the MAA synthesis in phytoplankton from remote Himalayan lakes. [ABSTRACT FROM AUTHOR]

The article presents a study which determined the light dependence of Photosystem II (PSII) photoinhibition and photorepair in 2 phylotypes of Symbiodinium displaying different susceptibilities to thermal stress. They say that the key mechanism minimising net photoinhibition in Strain A1 was photorepair. They examined the response of the tolerant strain in relation to the growth irradiance of the culture in order to evaluate the relative role of photoprotection and photorepair in counteracting photodamage as a function of the photoacclimation state of Symbiodinium.

The response of the coccolithophorid Emiliania huxleyi (Lohmann) W. H. Hay et H. Mohler to acute exposure to high photon flux densities (PFD) was examined in terms of PSII photoinhibition, photoprotection, and photorepair. The time and light dependencies of these processes were characterized as a function of the photoacclimation state of the alga. Low-light (LL) acclimated cells displayed a higher degree of photoinhibition, measured as decline in Fv /Fm, than high-light (HL) acclimated cells. However, HL cultures were more susceptible to photodamage but also more capable of compensating for it by performing a faster repair cycle. The relation between gross photoinhibition (observed in the presence of an inhibitor of repair) and PFD to which the algae were exposed deviated from linearity at high PFD, which calls into question the universality of current concepts of photoinhibition in mechanistic models. The light dependence of the de-epoxidation state (DPS) of the xanthophyll cycle (XC) pigments on the timescale of hours was the same in cells acclimated to LL and HL. However, HL cells were more efficient in realizing nonphotochemical quenching (NPQ) on short timescales, most likely due to a larger XC pool. LL cells displayed an increase in the PSII effective cross-section (σPSII) as a result of photoinhibition, which was observed also in HL cells when net photoinhibition was induced by blocking the D1 repair cycle. The link between σPSII and photoinhibition suggests that the population of PSII reaction centers (RCIIs) of E. huxleyi shares a common antenna, according to a “lake” organization of the light-harvesting complex. [ABSTRACT FROM AUTHOR]

Abstract: The consequences of UVB and UVA irradiation on hatch rate, mortality, and malformation were studied in embryonic zebrafish (Danio rerio). The use of zebrafish embryos has expanded from traditional developmental models to diverse studies, including many techniques utilizing light exposure. To characterize useful indicators of photodamage, the responses and threshold limits of UV radiation as a function of embryonic stage and fish source were evaluated. Significant differences in UVB susceptibility were observed in embryos at 3, 6–7, 12, and 24h post-fertilization (hpf), with the 1000-cell stage (3hpf) having greatest tolerance to UVB. Embryos derived from zebrafish raised in outdoor ponds were more tolerant to UVB than were embryos from laboratory-raised fish. Combinations of UVB and UVA exposure were used to confirm the presence of a competent photorepair system in zebrafish that could return otherwise malformed embryos to a normal phenotype. Overall, embryonic zebrafish had large tolerances (LD50 of 850J/cm2) to UVA, confirming their suitability for photoactivation and photorepair studies. [Copyright &y& Elsevier]

To determine if an Antarctic species repairs DNA at rates equivalent to warmer water equivalents, we examined repair of UV-damaged DNA in echinoid embryos and larvae. DNA repair by photoreactivation was compared in three species Sterechinus neumayeri (Antarctica), Evechinus chloroticus (New Zealand) and Diadema setosum (Tropical Australia) spanning a latitudinal gradient from polar (77.86°S) to tropical (19.25°S) environments. We compared rates of photoreactivation as a function of ambient and experimental temperature in all three species, and rates of photoreactivation as a function of embryonic developmental stage in Sterechinus. DNA damage was quantified from cyclobutane pyrimidine dimer (CPD) concentrations and rates of abnormal embryonic development. This study established that in the three species and in three developmental stages of Sterechinus, photoreactivation was the primary means of removing CPDs, was effective in repairing all CPDs in less than 24 h, and promoted significantly higher rates of normal development in UV-exposed embryos. CPD photorepair rate constant (k) in echinoid embryos ranged from 0.33 to 1.25 h-1, equating to a time to 50% repair of between 0.6 and 2.1 h and time to 90%repair between 3.6 and 13.6 h. We observed that experimental temperature influenced photoreactivation rate. In Diadema plutei, the photoreactivation rate constant increased from k=0.58 h-1 to 1.25 h-1, with a Q10=2.15 between 22°C and 32°C. When compared among the three species across experimental temperatures (-1.9 to 32°C), photoreactivation rates vary with a Q10=1.39. Photoreactivation rates were examined in three developmental stages of Sterechinus embryos, and while not significantly different, repair rates tended to be higher in the younger blastula and gastrula stages compared with later stage embryos. We concluded that photoreactivation is active in the Antarctic Sterechinus, but at a significantly slower (non-temperature compensated) rate. The low level of temperature compensation in photoreactivation may be one explanation for the relatively high sensitivity of Antarctic embryos to UV-R in comparison with non-Antarctic equivalents. [ABSTRACT FROM AUTHOR]

• Background Depletion of the stratospheric ozone layer leads to an increase in ultraviolet-B (UVB: 280–320 nm) radiation reaching the earth's surface, and the enhanced solar UVB radiation predicted by atmospheric models will result in reduction of growth and yield of crops in the future. Over the last two decades, extensive studies of the physiological, biochemical and morphological effects of UVB in plants, as well as the mechanisms of UVB resistance, have been carried out.• Scope In this review, we describe recent research into the mechanisms of UVB resistance in higher plants, with an emphasis on rice (Oryza sativa), one of the world's most important staple food crops. Recent studies have brought to light the following remarkable findings. UV-absorbing compounds accumulating in the epidermal cell layers have traditionally been considered to function as UV filters, and to play an important role in countering the damaging effects of UVB radiation. Although these compounds are effective in reducing cyclobutane pyrimidine dimer (CPD) induction in plants exposed to a challenge exposure to UVB, certain levels of CPD are maintained constitutively in light conditions containing UVB, regardless of the quantity or presence of visible light. These findings imply that the systems for repairing DNA damage and scavenging reactive oxygen species (ROS) are essential for plants to grow in light conditions containing UVB.• Conclusion CPD photolyase activity is a crucial factor determining the differences in UVB sensitivity between rice cultivars. The substitution of one or two bases in the CPD photolyase gene can alter the activity of the enzyme, and the associated resistance of the plant to UVB radiation. These findings open up the possibility, in the near future, of increasing the resistance of rice to UVB radiation, by selective breeding or bioengineering of the genes encoding CPD photolyase. [ABSTRACT FROM AUTHOR]

Sensitivity to ultraviolet-B (UVB) radiation (280–320 nm) varies widely among rice cultivars. We previously indicated that UV-resistant rice cultivars are better able to repair cyclobutane pyrimidine dimers (CPDs) through photorepair than are UV-sensitive cultivars. In this paper, we report that UVB sensitivity in rice, in part, is the result of defective CPD photolyase alleles. Surjamkhi ( indica) exhibited greater sensitivity to UVB radiation and was more deficient in CPD photorepair ability compared with UV-resistant Sasanishiki ( japonica). The deficiency in CPD photorepair in Surjamkhi resulted from changes in two nucleotides at positions 377 and 888 in the photolyase gene, causing alterations of two deduced amino acids at positions 126 and 296 in the photolyase enzyme. A linkage analysis in populations derived from Surjamkhi and Sasanishiki showed that UVB sensitivity is a quantitative inherited trait and that the CPD photolyase locus is tightly linked with a quantitative trait locus that explains a major portion of the genetic variation for this trait. These results suggest that spontaneously occurring mutations in the CPD photolyase gene cause different degrees of sensitivity to UVB in rice, and that the resistance of rice to UVB radiation could be increased by increasing the photolyase function through conventional breeding or bioengineering. [ABSTRACT FROM AUTHOR]

Cyclobutane pyrimidine dimers (CPDs) are the most frequent and deleterious lesions generated in the mammalian genome after UV-C irradiation. The persistence of these lesions in DNA can be toxic and mutagenic, and also represents a specific signal to apoptosis. To investigate the CPDs repair in situ and consequent UV-induced apoptosis in human cells, we generated a recombinant adenovirus vector containing the gene encoding a CPD-photolyase-EGFP fusion protein (Adphr-EGFP). Adphr-EGFP-infected cells are proficient in photorepair, which prevents apoptotic cell death in comparison with samples kept in the dark, indicating that the fusion protein is functional in CPD recognition and removal. By using local UV irradiation, foci of the photolyase fusion protein were observed in UV-damaged areas of the nuclei in colocalization with NER enzymes. Phr-EGFP migration to CPD sites and redistribution after photorepair was followed, and shown to present similar kinetics in normal or DNA-repair-deficient cells. To our knowledge, this is the first report of an investigation of CPDs repair in situ employing a CPD-photolyase-EGFP enzyme. The Adphr-EGFP vector can be an informative tool to investigate the repair and cellular consequences of UV-induced lesions in primary human cells. [ABSTRACT FROM AUTHOR]

Goldfish Carassius auratus embryos were subjected to artificial ultraviolet-B (UVB) radiation (280–320 nm) at various times during development to evaluate the effects on production of anatomically normal larvae. The UVB radiation used in these experiments included a higher proportion of shorter wavelengths compared to the natural spectrum. The development of embryos exposed to UVB for 2 or 4 h at 26 h post-fertilization was severely impaired whereas similar exposures at 50 or 74 h post-fertilization had no effect. A 2 h exposure to UVB commencing at 2 h post-fertilization did not adversely affect embryonic development whereas a 4 h exposure to a lower dose did. At 50 h post-fertilization, when embryos were normally resistant to UVB, denial of access to visible light and UVA before, during and after exposure to UVB caused impairment of development. Analysis of DNA fragment length after incubation with an endonuclease suggested that UVB damage at 50 h was caused by formation of pyrimidine dimers. This study demonstrated that the sensitivity of goldfish embryos to UVB varied during development and that resistance to UVB in later developmental stages included a photorepair mechanism. [ABSTRACT FROM AUTHOR]

Complementary DNA encoding a putative cyclobutane pyrimidine dimer (CPD)-specific DNA photolyase (CPD photolyase) was isolated from cucumber leaves. The deduced amino acid sequence of the cDNA exhibited high similarity to that of the Arabidopsis CPD photolyase. Transformation with the cDNA restored the impaired photorepair activity of an Escherichia coli mutant, indicating that this cDNA encodes a functional cucumber CPD photolyase (CsPHR). The level of CsPHR transcripts estimated by quantitative RT-PCR as well as the CPD photorepair activity in the cucumber first leaves showed diurnal changes, peaking at 09 : 00 and 12 : 00, respectively. Supplemental UV-B irradiation in the middle of the light period had little effect on the growth of the first leaves, while the supplemental irradiation in the early morning or late afternoon strongly retarded the leaf growth. These results suggest that the diurnal change in CPD photorepair activity, which is presumably regulated by the transcript level of CsPHR, may play an important role in minimizing the growth inhibition due to UV-B irradiation. [ABSTRACT FROM PUBLISHER]

I was trained as a physicist in graduate school. Hence, when I decided to go into the field of biophysics, it was natural that I concentrated on the effects of light on relatively simple biological systems, such as proteins. The wavelengths absorbed by the amino acid subunits of proteins are in the ultraviolet (UV). The wavelengths that affect the biological activities, the action spectra, also are in the UV, but are not necessarily parallel to the absorption spectra. Understanding these differences led me to investigate the action spectra for affecting nucleic acids, and the effects of UV on viruses and cells. The latter studies led me to the discovery of the important molecular nature of the damages affecting DNA (cyclobutane pyrimidine dimers) and to the discovery of nucleotide excision repair. Individuals with the genetic disease xeroderma pigmentosum (XP) are extraordinarily sensitive to sunlight-induced skin cancer. The finding, by James Cleaver, that their skin cells were defective in DNA repair strongly suggested that DNA damage was a key step in carcinogenesis. Such information was important for estimating the wavelengths in sunlight responsible for human skin cancer and for predicting the effects of ozone depletion on the incidence of non-melanoma skin cancer. It took experiments with backcross hybrid fish to call attention to the probable role of the longer UV wavelengths not absorbed by DNA in the induction of melanoma. These reflections trace the biophysicist’s path from molecules to melanoma. [Copyright &y& Elsevier]

Two photoproducts of DNA damage, i.e. cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs), induced by UV-B radiation in suspension-cultured tobacco cells were quantified by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. CPDs and 6-4PPs were induced in tobacco cells by UV-B radiation. Photorepair of CPDs was faster than that of 6-4PPs. UV-B radiation induces formation of CPDs and 6-4PPs even at 0 °C, but low temperature significantly decreases the UV-B-induced (in contrast to UV-C-induced) formation of CPDs and 6-4PPs. Low temperature also retarded the removal of CPDs and 6-4PPs under white light, and almost no photorepair of CPDs and 6-4PPs was detected at 0 °C. When purified DNA from tobacco cells grown in darkness was irradiated with UV-B, formation of CPDs and 6-4PPs took place at the same speed at different temperatures. It indicated that formation of CPDs and 6-4PPs induced by UV-B was temperature-independent in a non-cellular system. Based on our results for suspension-cultured tobacco cells, not only the photorepair but also UV-B-induced formation of CPDs and 6-4PPs are temperature-dependent. [Copyright &y& Elsevier]

Soybean [Glycine max (L.) Merr. cv. Hardee] and wheat (Triticum aestivum L. cv. Jori) were grown from seed under four ultraviolet-B irradiances and four levels of photosynthetically active radiation in a factorial design. The effects of ultraviolet-B radiation on leaf number and area, total dry matter production, dry weight of component organs, and plant height were compared between soybean and wheat. Ultraviolet-B radiation effects were dependent upon the level of photosynthetically active radiation incident during growth. Wheat and soybeans were both affected by low ultraviolet-B radiation flux densities; however, they differed markedly in their growth responses and biomass allocation patterns. Substantial interactions between ultraviolet-B and photosynthetically active radiation indicate a need for the measurement of longer wavelength radiation when evaluating the effects of ultraviolet-B radiation on plant growth in natural conditions. [ABSTRACT FROM AUTHOR]