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Nucleic Acid Photolysis by UV254 and the Impact of Virus Encapsidation
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2018-08-24 , DOI: 10.1021/acs.est.8b02308 Zhong Qiao 1 , Yinyin Ye 1 , Pin Hsuan Chang 1 , Devibaghya Thirunarayanan 1 , Krista R. Wigginton 1
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2018-08-24 , DOI: 10.1021/acs.est.8b02308 Zhong Qiao 1 , Yinyin Ye 1 , Pin Hsuan Chang 1 , Devibaghya Thirunarayanan 1 , Krista R. Wigginton 1
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Determining the influence of higher order structure on UVC photolysis will help inform predictions of nucleic acid fate and microorganism inactivation. We measured the direct UV254 photolysis kinetics of four model viral genomes composed of single-stranded and double-stranded RNA (ssRNA and dsRNA, respectively), as well as single-stranded and double-stranded DNA (ssDNA and dsDNA, respectively), in ultrapure water, in phosphate buffered saline (PBS), and encapsidated in their native virus particles. The photolysis rate constants of naked nucleic acids measured by qPCR (RT-qPCR for RNA) and normalized by the number of bases measured in a particular sequence exhibited the following trend: ssDNA > ssRNA ≈ dsDNA > dsRNA. In PBS, naked ssRNA bases reacted, on average, 24× faster than the dsRNA bases, whereas naked ssDNA bases reacted 4.3× faster than dsDNA bases. Endogenous indirect photolysis involving 1O2 and ·OH was ruled out as a major contributing factor in the reactions. A comparison of our measured rate constants with rate constants reported in the literature shows a general agreement among the nucleic acid UV254 direct photolysis kinetics. Our results underscore the high resistance of dsRNA to UVC photolysis and demonstrate the role that nucleic acid structure and solution chemistry play in photoreactivity.
中文翻译：
UV 254对核酸的光解作用和病毒衣壳化的影响
确定高阶结构对UVC光解的影响将有助于预测核酸命运和微生物失活的预测。我们测量了直接紫外线254在超纯水中的磷酸盐中的四个模型病毒基因组的光解动力学，这些病毒基因组由单链和双链RNA（分别为ssRNA和dsRNA）以及单链和双链DNA（分别为ssDNA和dsDNA）组成缓冲盐水（PBS），并封装在其天然病毒颗粒中。通过qPCR（针对RNA的RT-qPCR）测量并通过特定序列中测量的碱基数进行归一化的裸核酸的光解速率常数呈现以下趋势：ssDNA> ssRNA≈dsDNA> dsRNA。在PBS中，裸露的ssRNA碱基的反应平均速度比dsRNA碱基快24倍，而裸露的ssDNA碱基的反应速率则比dsDNA碱基快4.3倍。涉及1 O 2的内源性间接光解·OH被认为是反应中的主要贡献因素。将我们测得的速率常数与文献中报道的速率常数进行比较，结果表明，核酸UV 254直接光解动力学具有普遍的一致性。我们的结果强调了dsRNA对UVC光解的高抗性，并证明了核酸结构和溶液化学在光反应性中的作用。
更新日期：2018-08-26
中文翻译：
UV 254对核酸的光解作用和病毒衣壳化的影响
确定高阶结构对UVC光解的影响将有助于预测核酸命运和微生物失活的预测。我们测量了直接紫外线254在超纯水中的磷酸盐中的四个模型病毒基因组的光解动力学，这些病毒基因组由单链和双链RNA（分别为ssRNA和dsRNA）以及单链和双链DNA（分别为ssDNA和dsDNA）组成缓冲盐水（PBS），并封装在其天然病毒颗粒中。通过qPCR（针对RNA的RT-qPCR）测量并通过特定序列中测量的碱基数进行归一化的裸核酸的光解速率常数呈现以下趋势：ssDNA> ssRNA≈dsDNA> dsRNA。在PBS中，裸露的ssRNA碱基的反应平均速度比dsRNA碱基快24倍，而裸露的ssDNA碱基的反应速率则比dsDNA碱基快4.3倍。涉及1 O 2的内源性间接光解·OH被认为是反应中的主要贡献因素。将我们测得的速率常数与文献中报道的速率常数进行比较，结果表明，核酸UV 254直接光解动力学具有普遍的一致性。我们的结果强调了dsRNA对UVC光解的高抗性，并证明了核酸结构和溶液化学在光反应性中的作用。
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