The significance of the Dewar valence photoisomer as a UV radiation-induced DNA photoproduct in marine microbial communities.

Induction of pyrimidine dimers in DNA by solar UV radiation has drastic effects on microorganisms. To better define the nature of these DNA photoproducts in marine bacterioplankton and eukaryotes, a study was performed during a cruise along a latitudinal transect in the Pacific Ocean. The frequency of all possible cyclobutane pyrimidine dimers, pyrimidine (6-4) pyrimidone photoproducts (64PPs) and their related Dewar valence isomers (DEWs) was determined by high-performance liquid chromatography-mass spectrometry. Studied samples were bacterioplankton and eukaryotic fractions isolated from sea water either collected before sunrise or exposed to ambient sunlight from sunrise to sunset. Isolated DNA dosimeters were also exposed to daily sunlight for comparison purposes. A first major result was the observation in all samples of large amounts of DEWs, a class of photoproducts rarely considered outside photochemical studies. Evidence was obtained for a major role of UVA in the formation of these photoisomerization products of 64PPs. Considerations on the ratio between the different classes of photoproducts in basal and induced DNA damage suggests that photoenzymatic repair (PER) is an important DNA repair mechanism used by marine microorganisms occupying surface seawater in the open ocean. This result emphasizes the biological role of DEWs which are very poor substrate for PER.

Closed magnetic topology in the Venusian magnetotail and ion escape at Venus.

Venus, lacking an intrinsic global dipole magnetic field, serves as a textbook example of an induced magnetosphere, formed by interplanetary magnetic fields (IMF) enveloping the planet. Yet, various aspects of its magnetospheric dynamics and planetary ion outflows are complex and not well understood. Here we analyze plasma and magnetic field data acquired during the fourth Venus flyby of the Parker Solar Probe (PSP) mission and show evidence for closed topology in the nightside and downstream portion of the Venus magnetosphere (i.e., the magnetotail). The formation of the closed topology involves magnetic reconnection-a process rarely observed at non-magnetized planets. In addition, our study provides an evidence linking the cold Venusian ion flow in the magnetotail directly to magnetic connectivity to the ionosphere, akin to observations at Mars. These findings not only help the understanding of the complex ion flow patterns at Venus but also suggest that magnetic topology is one piece of key information for resolving ion escape mechanisms and thus the atmospheric evolution across various planetary environments and exoplanets.

GCN5 and E2F1 stimulate nucleotide excision repair by promoting H3K9 acetylation at sites of damage.

Chromatin structure is known to be a barrier to DNA repair and a large number of studies have now identified various factors that modify histones and remodel nucleosomes to facilitate repair. In response to ultraviolet (UV) radiation several histones are acetylated and this enhances the repair of DNA photoproducts by the nucleotide excision repair (NER) pathway. However, the molecular mechanism by which UV radiation induces histone acetylation to allow for efficient NER is not completely understood. We recently discovered that the E2F1 transcription factor accumulates at sites of UV-induced DNA damage and directly stimulates NER through a non-transcriptional mechanism. Here we demonstrate that E2F1 associates with the GCN5 acetyltransferase in response to UV radiation and recruits GCN5 to sites of damage. UV radiation induces the acetylation of histone H3 lysine 9 (H3K9) and this requires both GCN5 and E2F1. Moreover, as previously observed for E2F1, knock down of GCN5 results in impaired recruitment of NER factors to sites of damage and inefficient DNA repair. These findings demonstrate a direct role for GCN5 and E2F1 in NER involving H3K9 acetylation and increased accessibility to the NER machinery.

Ultraviolet A does not induce melanomas in a Xiphophorus hybrid fish model.

We examined the wavelength dependence of ultraviolet (UV) ra-diation (UVR)-induced melanoma in a Xiphophorus backcross hybrid model previously reported to be susceptible to melanoma induction by ultraviolet A (UVA) and visible light. Whereas ultraviolet B (UVB) irradiation of neonates yielded high frequencies of melanomas in pigmented fish, UVA irradiation resulted in melanoma frequencies that were not significantly different from unirradiated fish. Spontaneous and UV-induced melanoma frequencies correlated with the degree of pigmentation as expected from previous studies, and the histopathology phenotypes of the melanomas were not found in significantly different proportions in UV-treated and -untreated tumor-bearing fish. Our results support the conclusion that a brief early-life exposure to UVB radiation causes melanoma formation in this animal model. These data are consistent with an essential role for direct DNA damage, including cyclobutane dimers and (6-4) photoproducts, in the etiology of melanoma.

E2F1 localizes to sites of UV-induced DNA damage to enhance nucleotide excision repair.

The E2F1 transcription factor is a well known regulator of cell proliferation and apoptosis, but its role in the DNA damage response is less clear. Using a local UV irradiation technique and immunofluorescence staining, E2F1 is shown to accumulate at sites of DNA damage. Localization of E2F1 to UV-damaged DNA requires the ATM and Rad3-related (ATR) kinase and serine 31 of E2F1 but not an intact DNA binding domain. E2F1 deficiency does not appear to affect the expression of nucleotide excision repair (NER) factors, such as XPC and XPA. However, E2F1 depletion does impair the recruitment of NER factors to sites of damage and reduces the efficiency of DNA repair. E2F1 mutants unable to bind DNA or activate transcription retain the ability to stimulate NER. These findings demonstrate that E2F1 has a direct, non-transcriptional role in DNA repair involving increased recruitment of NER factors to sites of damage.

High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage.

High mobility group protein B1 (HMGB1) is a multifunctional protein with roles in chromatin structure, transcriptional regulation, V(D)J recombination, and inflammation. HMGB1 also binds to and bends damaged DNA, but the biological consequence of this interaction is not clearly understood. We have shown previously that HMGB1 binds cooperatively with nucleotide excision repair damage recognition proteins to triplex-directed psoralen DNA interstrand cross-links (ICLs). Thus, we hypothesized that HMGB1 modulates the repair of DNA damage in mammalian cells. We demonstrate here that mammalian cells lacking HMGB1 are hypersensitive to DNA damage induced by psoralen plus UVA irradiation (PUVA) or UVC radiation, showing less survival and increased mutagenesis. In addition, nucleotide excision repair efficiency is significantly decreased in the absence of HMGB1 as assessed by the repair and removal of UVC lesions from genomic DNA. We also explored the role of HMGB1 in chromatin remodeling upon DNA damage. Immunoblotting demonstrated that, in contrast to HMGB1 proficient cells, cells lacking HMGB1 showed no histone acetylation upon DNA damage. Additionally, purified HMGB1 protein enhanced chromatin formation in an in vitro chromatin assembly system. These results reveal a role for HMGB1 in the error-free repair of DNA lesions. Its absence leads to increased mutagenesis, decreased cell survival, and altered chromatin reorganization after DNA damage. Because strategies targeting HMGB1 are currently in development for treatment of sepsis and rheumatoid arthritis, our findings draw attention to potential adverse side effects of anti-HMGB1 therapy in patients with inflammatory diseases.

Ultraviolet radiation affects invasibility of lake ecosystems by warm-water fish.

Predicting where species invasions will occur remains a substantial challenge in ecology, but identifying factors that ultimately constrain the distribution of potential invaders could facilitate successful prediction. Whereas ultraviolet radiation (UVR) is recognized as an important factor controlling species distribution and community composition, the role of UVR in a habitat invasibility context has not been explored. Here we examine how underwater UVR can regulate warm-water fish invasion. In Lake Tahoe, California and Nevada, USA, established populations of exotic bluegill sunfish (Lepomis macrochirus) are currently limited to turbid, low-UVR embayments. An in situ incubation experiment that manipulated incident UVR exposure of larval bluegill, combined with an assessment of UVR exposure levels in nearshore habitats around Lake Tahoe, demonstrates that UVR can mediate habitat invasibility. Our findings suggest that the susceptibility to invasion by UVR sensitive species may increase in transparent aquatic systems threatened by declining water quality, and they highlight the importance of abiotic factors as regulators of invasion risk in ecosystems.

Direct effects of UV-B radiation on the freshwater heterotrophic nanoflagellate Paraphysomonas sp.

The formation of DNA photoproducts in organisms exposed to ambient levels of UV-B radiation can lead to death and/or reduced population growth in aquatic systems. Dependence on photoenzymatic repair to reverse DNA damage caused by UV-B radiation is demonstrated for Paraphysomonas sp., a member of a widely distributed genus of heterotrophic nanoflagellates. At 20 degrees C, Paraphysomonas sp. was exposed to a range of UV-B intensities encountered in natural systems. Populations of the flagellate survived and grew in a dose-dependent manner, but only when simultaneously exposed to photorepair radiation (PRR). In contrast, flagellates exposed to UV-B at 15 degrees C suffered 100% mortality except at the lowest UV-B level (with PRR) tested, which suggested a photorepair temperature optimum above 15 degrees C. After acute UV-B exposures, DNA damage (measured as the formation of pyrimidine dimers) was reduced only in organisms that underwent subsequent exposure to PRR. Populations kept in the dark after UV-B exposure maintained the initial levels of pyrimidine dimers. These results are the first to demonstrate the reliance of a heterotrophic flagellate on photoenzymatic DNA repair for survival from UV-B exposure.

Temperature effects on survival and DNA repair in four freshwater cladoceran Daphnia species exposed to UV radiation.

The biological responses of four freshwater daphniid species, Daphnia middendorffiana, D. pulicaria, D. pulex and D. parvula, to a single acute dose of ultraviolet B radiation (UVB) were compared. In addition to survival, we compared the induction of DNA damage (i.e. cyclobutane pyrimidine dimers) between species as well as the ability to repair this damage in the presence or absence of photoreactivating light. All four species showed high levels of shielding against DNA damage when compared to damage induced in purified DNA dosimeters at the same time and dose. Significant variation in survival was observed between species depending on temperature and light conditions. Contrary to our expectations, all species showed significantly higher survival and light-dependent DNA damage removal rates at 10 degrees C compared to 20 degrees C, suggesting that the enhanced rate of photoenzymatic repair (PER) at the lower temperature contributed significantly to the recovery of these organisms from UVB. PER was highly effective in promoting survival of three of the four species at 10 degrees C, but at 20 degrees C it was only partially effective in two species, and ineffective in two others. None of the species showed significant dark repair at 20 degrees C and only D. pulicaria showed a significant capacity at 10 degrees C. Two species, D. middendorffiana and D. pulex, showed some short-term survival at 10 degrees C in absence of PER despite their inability to repair any appreciable amount of DNA damage in the dark. All species died rapidly at 20 degrees C in absence of PER, as predicted from complete or near-absence of nucleotide excision repair (NER). Overall, the protective effects of tissue structure and pigmentation were similar in all Daphnia species tested and greatly mitigated the absorption of UVB by DNA and its damaging effects. Surprisingly, the visibly melanotic D. middendorffiana was not better shielded from DNA damage than the three non-melanotic species, and in fact suffered the highest damage rates. Melanin content in this species was not temperature dependent under the experimental growth conditions, and so did not contribute to temperature-dependent responses. It is evident that different species within the same genus have developed diverse biological responses to UVB. Our data strongly suggest that DNA damage is lethal to Daphnia and that photoenzymatic repair is the primary mechanism for removing these lesions. In the absence of light, few species are capable of removing any DNA damage. Surprisingly, the single species in which significant excision repair was detected did so only at reduced temperature. This temperature-dependence of excision repair is striking and may reflect adaptations of certain organisms to stress in a complex and changing environment.

Sunlight-induced DNA damage in marine micro-organisms collected along a latitudinal gradient from 70 degrees N to 68 degrees S.

We examined ultraviolet radiation (UVR)-induced DNA damage in marine micro-organisms collected from surface seawater along a latitudinal transect in the Central Pacific Ocean from 70 degrees N to 68 degrees S. Samples were collected predawn and incubated under ambient UVR in transparent incubators at in situ temperatures until late afternoon at which time they were filtered into primarily bacterioplankton and eukaryotic fractions. Cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts [(6-4)PDs] were quantified in DNA extracts using radioimmunoassays. UVB was lowest in the polar regions and highest near the equator and correlations between UVB and DNA damage were observed. The eukaryotic fraction showed significant CPDs across the entire transect; (6-4)PDs were detected only in the tropics. The bacterial fraction showed no accumulation of (6-4)PDs at any latitude, although residual (6-4)PDs were observed. Bacterial cell volumes were greatest in the sub-Arctic and northern temperate latitudes and lower in the tropics and southern hemisphere, a unique observation that parallels Bergmann's rule. A strong negative correlation was observed between cell volume and CPDs. The environmental impact of solar UVR on marine micro-organisms in the open ocean is complex and our results suggest that several factors such as DNA repair, cell size, temperature, salinity, nutrients and species composition are important in determining relative sensitivity.

Mapping the Lunar Wake Potential Structure With ARTEMIS Data.

The refilling of the lunar wake is relatively well explained by the theory of 1-D plasma expansion into a vacuum; however, the field-aligned wake potential is not a directly measured quantity, and thus, a statistical analysis of wake potentials at high altitudes has not been previously performed. In this study, we obtain the wake potential by comparing the field-aligned electron distributions inside and outside of the lunar wake measured by the two probes of the Acceleration, Reconnection, Turbulence, and Electrodynamics of Moon's Interaction with the Sun (ARTEMIS) mission. The derived potentials from ARTEMIS data vary with solar wind electron temperature and bulk flow velocity as the theory predicts. We also expand the 1-D plasma theory to 2-D in the plane of the interplanetary magnetic field and the solar wind velocity to examine how a tilted interplanetary magnetic field affects the wake potential structure. As the expansion time for the two sides of the wake differs, a wake potential asymmetry is developed in our model. This asymmetry is confirmed by the data-derived wake potentials. Moreover, ambipolar electric fields are obtained from both the modeled and data-derived wake potentials and show good agreement. Lastly, we examine the effects of the solar wind strahl-electron population on the wake potential structure, which appears to cause a net potential difference across the lunar shadow. This may imply that the disturbance of the wake plasma expansion extends farther outside the wake than previous plasma-expansion theories have predicted.

CALIPSO (IIR-CALIOP) Retrievals of Cirrus Cloud Ice Particle Concentrations.

A new satellite remote sensing method is described whereby the sensitivity of thermal infrared wave resonance absorption to small ice crystals is exploited to estimate cirrus cloud ice particle number concentration N, effective diameter De, and ice water content IWC. This method uses co-located observations from the Infrared Imaging Radiometer (IIR) and from the CALIOP (Cloud and Aerosol Lidar with Orthogonal Polarization) lidar aboard the CALIPSO (Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation) polar orbiting satellite, employing IIR channels at 10.6 µm and 12.05 µm. Using particle size distributions measured over several flights of the TC4 (Tropical Composition, Cloud and Climate Coupling) and the mid-latitudes SPARTICUS (Small Particles in Cirrus) field campaigns, we show for the first time that N/IWC is tightly related to ßeff; the ratio of effective absorption optical depths at 12.05 µm and 10.6 µm. Relationships developed from in situ aircraft measurements are applied to ßeff derived from IIR measurements to retrieve N. This satellite remote sensing method is constrained by measurements of ßeff from the IIR and is by essence sensitive to the smallest ice crystals. Retrieval uncertainties are discussed, including uncertainties related to in situ measurement of small ice crystals (D < 15 µm), which are studied through comparisons with IIR ßeff. The method is applied here to single-layered semi-transparent clouds having a visible optical depth between about 0.3 and 3, where cloud base temperature is ≤ 235 K. Two years of CALIPSO data have been analyzed for the years 2008 and 2013, with the dependence of cirrus cloud N and De on altitude, temperature, latitude, season (winter vs. summer) and topography (land vs. ocean) described. The results for the mid-latitudes show a considerable dependence on season. In the high latitudes, N tends to be highest and De smallest, whereas the opposite is true for the tropics. The frequency of occurrence of these relatively thick cirrus clouds exhibited a strong seasonal dependence in the high latitudes, with the occurrence frequency during Arctic winter being at least twice that of any other season. Processes that could potentially explain some of these micro-and macroscopic cloud phenomena are discussed.

Anthropogenic Aerosol Indirect Effects in Cirrus Clouds.

We have implemented a parameterization for forming ice in large-scale cirrus clouds that accounts for the changes in updrafts associated with a spectrum of waves acting within each time step in the model. This allows us to account for the frequency of homogeneous and heterogeneous freezing events that occur within each time step of the model and helps to determine more realistic ice number concentrations as well as changes to ice number concentrations. The model is able to fit observations of ice number at the lowest temperatures in the tropical tropopause but is still somewhat high in tropical latitudes with temperatures between 195°K and 215°K. The climate forcings associated with different representations of heterogeneous ice nuclei (IN or INPs) are primarily negative unless large additions of IN are made, such as when we assumed that all aircraft soot acts as an IN. However, they can be close to zero if it is assumed that all background dust can act as an INP irrespective of how much sulfate is deposited on these particles. Our best estimate for the forcing of anthropogenic aircraft soot in this model is -0.2 ± 0.06 W/m2, while that from anthropogenic fossil/biofuel soot is -0.093 ± 0.033 W/m2. Natural and anthropogenic open biomass burning leads to a net forcing of -0.057 ± 0.05 W/m2.

Crosslinks rather than strand breaks determine access to ancient DNA sequences from frozen sediments.

Diagenesis was studied in DNA obtained from Siberian permafrost (permanently frozen soil) ranging from 10,000 to 400,000 years in age. Despite optimal preservation conditions, we found the sedimentary DNA to be severely modified by interstrand crosslinks; single- and double-stranded breaks; and freely exposed sugar, phosphate, and hydroxyl groups. Intriguingly, interstrand crosslinks were found to accumulate approximately 100 times faster than single-stranded breaks, suggesting that crosslinking rather than depurination is the primary limiting factor for ancient DNA amplification under frozen conditions. The results question the reliability of the commonly used models relying on depurination kinetics for predicting the long-term survival of DNA under permafrost conditions and suggest that new strategies for repair of ancient DNA must be considered if the yield of amplifiable DNA from permafrost sediments is to be significantly increased. Using the obtained rate constant for interstrand crosslinks the maximal survival time of amplifiable 120-bp fragments of bacterial 16S ribosomal DNA was estimated to be approximately 400,000 years. Additionally, a clear relationship was found between DNA damage and sample age, contradicting previously raised concerns about the possible leaching of free DNA molecules between permafrost layers.

Regulation of epidermal apoptosis and DNA repair by E2F1 in response to ultraviolet B radiation.

The E2F1 transcription factor regulates the expression of genes involved in cell proliferation, apoptosis and DNA repair. Following DNA damage, E2F1 is phosphorylated and stabilized, but the physiological role of E2F1 in the response to DNA damage is unclear. We find that mice lacking E2F1 have increased levels of epidermal apoptosis compared to wild-type mice following exposure to ultraviolet B (UVB) radiation. Moreover, transgenic overexpression of E2F1 in basal layer keratinocytes suppresses apoptosis induced by UVB. Inhibition of UVB-induced apoptosis by E2F1 is unexpected given that most studies have demonstrated a proapoptotic function for E2F1. E2F1-mediated suppression of apoptosis does not involve alterations in mitogen-activated protein kinase activation or Bcl-2 downregulation in response to UVB and is independent of p53. Instead, inhibition of UVB-induced apoptosis by E2F1 correlates with a stimulation of DNA repair. Mice lacking E2F1 are impaired for the removal of DNA photoproducts, while E2F1 transgenic mice repair UVB-induced DNA damage at an accelerated rate compared to wild-type mice. These findings suggest that E2F1 participates in the response to UVB by promoting DNA repair and suppressing apoptosis.

Quantifying ultraviolet radiation mortality risk in bluegill larvae: effects of nest location.

Ultraviolet (UV) radiation (280-400 nm) is an increasing threat to aquatic organisms due to stratospheric ozone depletion and reductions in concentrations of dissolved organic carbon. Because fish are most vulnerable to UV during the egg and larval stages, parental spawning site selection can strongly influence mortality risk. We examined the role of nest location in determining UV-induced mortality risk for bluegill (Lepomis macrochirus) in Lake Giles, Pennsylvania, USA. In a series of five short-term incubation experiments, we found that survival of yolk sac larvae across the range of depths at which bluegill spawn was significantly lower in the presence of ambient-UV levels relative to larvae that were shielded from UV radiation. In addition, survival decreased as a function of cumulative UV exposure, as measured by the number of cyclobutane pyrimidine dimers per megabase DNA in DNA dosimeters. Although UV had the potential to significantly reduce larval survival, DNA dosimeters placed in bluegill nests concurrently with incubation experiments indicated that most nests were exposed to relatively low levels of UV. Only 19% of nests had predicted UV-induced mortality greater than 25%. Consequently, current levels of UV may be an important mortality source at the level of individual nests, but not at the population level. One reason for the weak predicted effect of UV on bluegill survival is that many nests were located at depths by which much of the incident UV had been attenuated. In addition, many of the shallower nests were protected by overhanging trees or other submerged structures. It is important to note that Lake Giles is highly transparent and therefore not representative of all lakes in which bluegill are found. Nevertheless, Lake Giles is a natural system and may be representative of north temperate lakes in the future.

Interspecific variation in UV defense mechanisms among temperate freshwater fishes.

An important step in predicting the effects of future increases in UV radiation (UVR) is to evaluate the mechanisms that organisms use to prevent and repair DNA damage and determine how those mechanisms influence UVR sensitivity. Damage is prevented to varying degrees through photoprotection and repaired via two main pathways: nucleotide excision repair and photoenzymatic repair. At present, little is known about the generality or similarity of these defenses among temperate freshwater fishes. We used laboratory experiments to compare UVR defense mechanisms among five freshwater fish species representing four families and three orders. Purified DNA, freeze-killed larvae and live larvae were exposed to UVB radiation for 12 h in the presence or absence of photorepair radiation. After exposure, we quantified frequencies of cyclobutane pyrimidine dimers in each exposure treatment. All five species used photoprotection and proportional decreases in dimer frequency were similar for all species. Evidence of excision repair was also found for all species but proportional decreases in photoproduct frequencies varied among species. Finally, evidence of photoenzymatic repair was found for only two of the five species.

Quantification of photoproducts in mammalian cell DNA using radioimmunoassay.

Over the past 20 yr, the use of polyclonal and monoclonal antibodies to quantify damage in DNA has burgeoned. Immunoassays offer distinct advantages over other anaytical procedures currently used to measure DNA damage including adaptability, sensitivity and selectivity. This combination of attributes allows for the development of powerful analytical techniques to visualize and quantify specific types of DNA damage in cells and organisms exposed to subtoxic levels of xenobiotics with distinct advantages over the other procedures in the analysis of DNA damage in human and environmental samples. Radioimmunoassay (RIA) is readily applied to a variety of biological materials and has typically been used to measure DNA damage in cell and organ cultures, tissue sections and biopsies, buccal cells, bone marrow aspirates, peripheral blood lymphocytes, and urine. Here we describe the use of a very sensitive RIA for the specific quantitation of cyclobutane dimers and (6-4) photoproducts in DNA extracted from mammalian cells and tissues.

Protein kinase C epsilon is an endogenous photosensitizer that enhances ultraviolet radiation-induced cutaneous damage and development of squamous cell carcinomas.

Chronic exposure to UV radiation (UVR), especially in the UVA (315-400 nm) and UVB (280-315 nm) spectrum of sunlight, is the major risk factor for the development of nonmelanoma skin cancer. UVR is a complete carcinogen, which both initiates and promotes carcinogenesis. We found that protein kinase C epsilon (PKCepsilon), a member of the phospholipid-dependent threonine/serine kinase family, is an endogenous photosensitizer, the overexpression of which in the epidermis increases the susceptibility of mice to UVR-induced cutaneous damage and development of squamous cell carcinoma. The PKCepsilon transgenic mouse (FVB/N) lines 224 and 215 overexpressed 8- and 18-fold PKCepsilon protein, respectively, over endogenous levels in basal epidermal cells. UVR exposure (1 kJ/m(2) three times weekly) induced irreparable skin damage in high PKCepsilon-overexpressing mouse line 215. However, the PKCepsilon transgenic mouse line 224, when exposed to UVR (2 kJ/m(2) three times weekly), exhibited minimum cutaneous damage but increased squamous cell carcinoma multiplicity by 3-fold and decreased tumor latency by 12 weeks. UVR exposure of PKCepsilon transgenic mice compared with wild-type littermates (1) elevated the levels of neither cyclobutane pyrimidine dimer nor pyrimidine (6-4) pyrimidone dimer, (2) reduced the appearance of sunburn cells, (3) induced extensive hyperplasia and increased the levels of mouse skin tumor promoter marker ornithine decarboxylase, and (4) elevated the levels of tumor necrosis factor alpha (TNFalpha) and other growth stimulatory cytokines, granulocyte colony-stimulating factor, and granulocyte macrophage colony-stimulating factor. The role of TNFalpha in UVR-induced cutaneous damage was evaluated using PKCepsilon transgenic mice deficient in TNFalpha. UVR treatment three times weekly for 13 weeks at 2 kJ/m(2) induced severe cutaneous damage in PKCepsilon transgenic mice (line 215), which was partially prevented in PKCepsilon-transgenic TNFalpha-knockout mice. Taken together, the results indicate that PKCepsilon signals UVR-induced TNFalpha release that is linked, at least in part, to the photosensitivity of PKCepsilon transgenic mice.

Effective intra-S checkpoint responses to UVC in primary human melanocytes and melanoma cell lines.

The objective of this study was to assess potential functional attenuation or inactivation of the intra-S checkpoint during melanoma development. Proliferating cultures of skin melanocytes, fibroblasts, and melanoma cell lines were exposed to increasing fluences of UVC and intra-S checkpoint responses were quantified. Melanocytes displayed stereotypic intra-S checkpoint responses to UVC qualitatively and quantitatively equivalent to those previously demonstrated in skin fibroblasts. In comparison with fibroblasts, primary melanocytes displayed reduced UVC-induced inhibition of DNA strand growth and enhanced degradation of p21Waf1 after UVC, suggestive of enhanced bypass of UVC-induced DNA photoproducts. All nine melanoma cell lines examined, including those with activating mutations in BRAF or NRAS oncogenes, also displayed proficiency in activation of the intra-S checkpoint in response to UVC irradiation. The results indicate that bypass of oncogene-induced senescence during melanoma development was not associated with inactivation of the intra-S checkpoint response to UVC-induced DNA replication stress.