UVA generates pyrimidine dimers in DNA directly.

Center for Biologically Inspired Materials and Material Systems, Duke University, Durham, North Carolina, USA.
Biophysical Journal (Impact Factor: 3.67). 03/2009; 96(3):1151-8. DOI: 10.1016/j.bpj.2008.10.030
Source: PubMed

ABSTRACT There is increasing evidence that UVA radiation, which makes up approximately 95% of the solar UV light reaching the Earth's surface and is also commonly used for cosmetic purposes, is genotoxic. However, in contrast to UVC and UVB, the mechanisms by which UVA produces various DNA lesions are still unclear. In addition, the relative amounts of various types of UVA lesions and their mutagenic significance are also a subject of debate. Here, we exploit atomic force microscopy (AFM) imaging of individual DNA molecules, alone and in complexes with a suite of DNA repair enzymes and antibodies, to directly quantify UVA damage and reexamine its basic mechanisms at a single-molecule level. By combining the activity of endonuclease IV and T4 endonuclease V on highly purified and UVA-irradiated pUC18 plasmids, we show by direct AFM imaging that UVA produces a significant amount of abasic sites and cyclobutane pyrimidine dimers (CPDs). However, we find that only approximately 60% of the T4 endonuclease V-sensitive sites, which are commonly counted as CPDs, are true CPDs; the other 40% are abasic sites. Most importantly, our results obtained by AFM imaging of highly purified native and synthetic DNA using T4 endonuclease V, photolyase, and anti-CPD antibodies strongly suggest that CPDs are produced by UVA directly. Thus, our observations contradict the predominant view that as-yet-unidentified photosensitizers are required to transfer the energy of UVA to DNA to produce CPDs. Our results may help to resolve the long-standing controversy about the origin of UVA-produced CPDs in DNA.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: This study compared biological responses of normal human fibroblasts (NHF1) to three sources of ultraviolet radiation (UVR), emitting UVC wavelengths, UVB wavelengths, or a combination of UVA and UVB (solar simulator; emission spectrum, 94.3% UVA and 5.7% UVB). The endpoints measured were cytotoxicity, intra-S checkpoint activation, inhibition of DNA replication, and mutagenicity. Results show that the magnitude of each response to the indicated radiation sources was best predicted by the density of DNA cyclobutane pyrimidine dimers (CPD). The density of 6-4 pyrimidine-pyrimidone photoproducts (6-4PP) was highest in DNA from UVC-irradiated cells (14% of CPD) as compared to those exposed to UVB (11%) or UVA-UVB (7%). The solar simulator source, under the experimental conditions described here, did not induce the formation of 8-oxo-7,8-dihydroguanine (8-oxo-dG) in NHF1 above background levels. Taken together, these results suggest that CPD play a dominant role in DNA damage responses and highlight the importance of using endogenous biomarkers to compare and report biological effects induced by different sources of UVR. This article is protected by copyright. All rights reserved.
    Photochemistry and Photobiology 10/2013; · 2.29 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Escherichia coli and enterococci have been previously reported to differ in the mechanisms and conditions that affect their sunlight-mediated inactivation in waste stabilization ponds. This study was undertaken to further characterize these mechanisms, using simulated sunlight and single strains of laboratory-grown E. coli and Enterococcus faecalis, with a focus on characterizing the contribution of exogenous reactive oxygen species to the inactivation process. We found that direct damage by UVB light (280-320 nm) was not a significant inactivation mechanism for either organism. E. coli inactivation was strongly dependent on dissolved oxygen concentrations and the presence of UVB wavelengths but E. coli were not susceptible to inactivation by exogenous sensitizers present in waste stabilization pond water. In contrast, E. faecalis inactivation in pond water occurred primarily through exogenous mechanisms, with strong evidence that singlet oxygen is an important transient reactive species. The exogenous mechanism could utilize wavelengths into the visible spectrum and sensitizers were mainly colloidal, distributed between 0.2 and ∼1 μm in size. Singlet oxygen is likely an important endogenous species in both E. faecalis and E. coli inactivation due to sunlight. Although the two organisms had similar inactivation rates in buffered, clear water, the inactivation rate of E. faecalis was 7 times greater than that of E. coli in air-saturated pond water at circumneutral pH due to its susceptibility to exogenous sensitizers and longer wavelengths.
    Water Research 10/2013; · 4.66 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The association between various measures of sun exposure and melanoma risk is quite complex to dissect as many case-control studies of melanoma included different subtypes of melanomas which are likely to be biologically different, so interpretation of the data is difficult. Screening bias in countries with high levels of sun exposure is also an issue. Now that progress is being made in the genetic subclassification of melanoma tumours, it is apparent that melanomas have different somatic changes according to body sites/histological subtypes and that UV exposure may be relevant for some but not all types of melanomas. Melanoma behaviour also points to non-sun-related risk factors, and complex gene-environment interactions are likely. As UV exposure is the only environmental factor ever linked to melanoma, it is still prudent to avoid excessive sun exposure and sunburn especially in poor tanners. However, the impact of strict sun avoidance, which should not be recommended, may take years to be apparent as vitamin D deficiency is a now a common health issue in Caucasian populations, with a significant impact on health in general.
    Current Oncology Reports 10/2013; · 3.33 Impact Factor

Full-text (2 Sources)

Available from
May 20, 2014