Comparative analysis of different laser systems to study cellular responses to DNA damage in mammalian cells.
Comparative analysis of different laser systems to study cellular responses to DNA damage in mammalian cells.
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DOI：
10.1093/nar/gkp221
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发表时间：
2009-05
影响因子：
14.9
通讯作者：
Berns MW
中科院分区：
文献类型：
--
作者：
Kong X;Mohanty SK;Stephens J;Heale JT;Gomez-Godinez V;Shi LZ;Kim JS;Yokomori K;Berns MW
正确识别和修复DNA损伤是细胞保护其基因组完整性的关键。利用波长从紫外线A （UVA）到近红外（NIR）的激光微照射，可以诱导细胞核内特定区域的损伤，是有效分析哺乳动物细胞内DNA双链断裂（DSB）损伤识别过程的一种创新技术。然而，不同激光系统的损伤诱导特性尚未得到充分的研究。本文比较了含溴脱氧尿苷（BrdU）和不含溴脱氧尿苷（BrdU）的纳秒态氮337 nm UVA激光、纳秒态和皮秒态532 nm绿色二次谐波Nd:YAG激光以及飞秒态近红外800 nm Ti：蓝宝石激光的损伤类型和相应的细胞反应。交联损伤（没有明显的核苷酸切除修复因子招募）和单链断裂（有相应的修复因子招募）在所有三个波长中都很常见。有趣的是，没有BrdU的UVA只会产生碱基损伤和异常的DSB反应。此外，阈值H2AX磷酸化诱导所需的总能量在不同的激光系统之间有所不同。结果表明，不同波长和脉冲持续时间对损伤机制的影响不同。讨论了各系统的优缺点。
Proper recognition and repair of DNA damage is critical for the cell to protect its genomic integrity. Laser microirradiation ranging in wavelength from ultraviolet A (UVA) to near-infrared (NIR) can be used to induce damage in a defined region in the cell nucleus, representing an innovative technology to effectively analyze the in vivo DNA double-strand break (DSB) damage recognition process in mammalian cells. However, the damage-inducing characteristics of the different laser systems have not been fully investigated. Here we compare the nanosecond nitrogen 337 nm UVA laser with and without bromodeoxyuridine (BrdU), the nanosecond and picosecond 532 nm green second-harmonic Nd:YAG, and the femtosecond NIR 800 nm Ti:sapphire laser with regard to the type(s) of damage and corresponding cellular responses. Crosslinking damage (without significant nucleotide excision repair factor recruitment) and single-strand breaks (with corresponding repair factor recruitment) were common among all three wavelengths. Interestingly, UVA without BrdU uniquely produced base damage and aberrant DSB responses. Furthermore, the total energy required for the threshold H2AX phosphorylation induction was found to vary between the individual laser systems. The results indicate the involvement of different damage mechanisms dictated by wavelength and pulse duration. The advantages and disadvantages of each system are discussed.
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影响因子：
11.4
作者：
Kubota, Y;Nash, RA;Lindahl, T
通讯作者：
Lindahl, T
DOI：
10.1083/jcb.200510130
发表时间：
2006-04-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bekker-Jensen S;Lukas C;Kitagawa R;Melander F;Kastan MB;Bartek J;Lukas J
通讯作者：
Lukas J
影响因子：
4.8
作者：
Kim, JS;Krasieva, TB;Yokomori, K
通讯作者：
Yokomori, K
影响因子：
3.4
作者：
Botvinick, EL;Venugopalan, V;Berns, MW
通讯作者：
Berns, MW
影响因子：
10.5
作者：
Kitagawa, R;Bakkenist, CJ;Kastan, MB
通讯作者：
Kastan, MB