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首页> 美国卫生研究院文献>The FASEB Journal >Wavelength dependence of ultraviolet radiation-induced DNA damage as determined by laser irradiation suggests that cyclobutane pyrimidine dimers are the principal DNA lesions produced by terrestrial sunlight
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Wavelength dependence of ultraviolet radiation-induced DNA damage as determined by laser irradiation suggests that cyclobutane pyrimidine dimers are the principal DNA lesions produced by terrestrial sunlight

机译：激光辐照确定的紫外线辐射引起的DNA损伤的波长依赖性表明环丁烷嘧啶二聚体是陆地阳光产生的主要DNA损伤

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To elucidate the involvement of specific ultraviolet (UV) wavelengths in solar mutagenesis, we used a laser system to investigate the induction of DNA damage, both in the overall genome and at the nucleotide resolution level, in the genomic DNA of transgenic Big Blue mouse fibroblasts irradiated with a series of UV wavelengths, inclusive of UVC (λ<280 nm), UVB (λ=280–320 nm), and UVA (λ>320 nm). Subsequently, we sought correlation between the locations of UV-induced DNA lesions in the cII transgene of irradiated DNA samples and the frequency distribution and codon position of the induced cII mutations in counterpart mouse cells irradiated with simulated sunlight. Using a combination of enzymatic digestion assays coupled with gel electrophoresis, immunodot blot assays, and DNA footprinting assays, we demonstrated a unique wavelength-dependent formation of photodimeric lesions, i.e., cyclobutane pyrimidine dimers (CPDs) and (6–4) photoproducts [(6–4)PPs], based on direct UV absorption of DNA, in irradiated mouse genomic DNA, which could partially explain the induction of mutations in mouse cells irradiated with simulated sunlight. Most notably, there was a divergence of CPD and (6–4)PP formation at an irradiation wavelength of 296 nm in mouse genomic DNA. Whereas substantial formation of (6–4)PPs was detectable in samples irradiated at this wavelength, which intensified as the irradiation wavelength decreased, only small quantities of these lesions were found in samples irradiated at wavelengths of 300–305 nm, with no detectable level of (6–4)PPs in samples irradiated with longer wavelengths. Although CPD formation followed the same pattern of increase with decreasing wavelengths of irradiation, there were substantial levels of CPDs in samples irradiated with UVB wavelengths borderlined with UVA, and small but detectable levels of these lesions in samples irradiated with longer wavelengths. Because the terrestrial sunlight spectrum rolls off sharply at wavelengths ∼300 nm, our findings suggest that CPDs are the principal lesion responsible for most DNA damage-dependent biological effects of sunlight.—Besaratinia, A., Yoon, J. -I., Schroeder, C., Bradforth, S. E., Cockburn, M., Pfeifer, G. P. Wavelength dependence of ultraviolet radiation-induced DNA damage as determined by laser irradiation suggests that cyclobutane pyrimidine dimers are the principal DNA lesions produced by terrestrial sunlight.
机译：为了阐明特定的紫外线（UV）波长在太阳诱变中的作用，我们使用激光系统研究了转基因大蓝小鼠成纤维细胞的基因组DNA在整个基因组和核苷酸分辨率水平对DNA损伤的诱导作用。用一系列UV波长照射，包括UVC（λ<280 nm），UVB（λ= 280–320 nm）和UVA（λ> 320 nm）。随后，我们寻求辐照的DNA样品的cII转基因中紫外线诱导的DNA损伤的位置与模拟太阳光辐照的配对小鼠细胞中诱导的cII突变的频率分布和密码子位置之间的相关性。通过结合酶促消化测定，凝胶电泳，免疫斑点印迹测定和DNA足迹测定，我们证明了光二聚体损伤的独特的波长依赖性形成，即环丁烷嘧啶二聚体（CPD）和（6–4）光产物[[（ 6–4）PPs]，是基于被辐照的小鼠基因组DNA中DNA的直接紫外线吸收，这可以部分解释诱导模拟阳光辐照的小鼠细胞中突变的产生。最值得注意的是，在小鼠基因组DNA的照射波长为296 nm时，CPD和（6–4）PP形成存在差异。尽管在此波长下照射的样品中可检测到大量的（6–4）PPs形成，并且随着照射波长的减小而加剧，但在300–305 nm波长下照射的样品中仅发现少量这些病变，没有检测到水平较长波长的样品中（6–4）PP的比例。尽管CPD的形成随着辐射波长的减少而遵循相同的增加模式，但是在以UVA为边界的UVB波长辐照的样品中，CPD的水平较高，而在以更长波长辐照的样品中，这些病变的水平很小但可检测。由于地球的日光光谱在约300 nm的波长处急剧下降，因此我们的发现表明CPD是造成大多数依赖DNA损伤的日光生物效应的主要病变。—Besaratinia，A.，Yoon，J. -I。，Schroeder ，C.，Bradforth，SE，Cockburn，M.，Pfeifer，GP通过激光照射确定的紫外线辐射引起的DNA损伤的波长依赖性表明，环丁烷嘧啶二聚体是陆地阳光产生的主要DNA损伤。

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