Mechanisms of Human Adenovirus Inactivation by Sunlight and UVC Light as Examined by Quantitative PCR and Quantitative Proteomics
Mechanisms of Human Adenovirus Inactivation by Sunlight and UVC Light as Examined by Quantitative PCR and Quantitative Proteomics
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DOI：
10.1128/aem.03457-12
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发表时间：
2013-02-01
影响因子：
4.4
通讯作者：
Kohn, Tamar
中科院分区：
文献类型：
--
作者：
Bosshard, Franziska;Armand, Florence;Kohn, Tamar
人类腺病毒（hav）是工业化国家和发展中国家的重要病原体。hav已被证明对单色UVC光具有相对的抵抗力。相比之下，多色UVC光是一种更有效的消毒手段，可能是由于病毒蛋白参与了失活机制。最后，我们对hav的太阳消毒了解甚少。本文阐述了紫外线光和直接、间接太阳消毒灭活hav的动力学和机理。PCR和质谱法用于量化基因组和蛋白质降解的程度，并定位hav蛋白中的受影响区域。为此，我们首次采用了人病毒细胞培养（SILAC）中氨基酸稳定同位素标记的方法。UVC光和全太阳光光谱的灭活被发现能有效地灭活hav，而uva -可见光仅在外部敏化剂（间接太阳消毒）存在的情况下引起灭活。紫外线对基因组的损害显著，但对日光消毒的影响较小。相比之下，间接日光消毒表现出广泛的蛋白质降解。特别是，纤维蛋白和纤维蛋白内负责宿主结合的氨基酸被证明会降解。此外，penton蛋白的中心结构域被破坏，这可能会抑制与纤维蛋白的相互作用，导致感染的初始阶段中断。然而，六邻体蛋白的损伤似乎只影响不直接参与感染周期的区域。
Human adenoviruses (HAdV) are important pathogens in both industrialized and developing nations. HAdV has been shown to be relatively resistant to monochromatic UVC light. Polychromatic UVC light, in contrast, is a more effective means of disinfection, presumably due to the involvement of viral proteins in the inactivation mechanism. Solar disinfection of HAdV, finally, is only poorly understood. In this paper, the kinetics and mechanism of HAdV inactivation by UVC light and direct and indirect solar disinfection are elucidated. PCR and mass spectrometry were employed to quantify the extent of genome and protein degradation and to localize the affected regions in the HAdV proteins. For this purpose, we used for the first time an approach involving stable isotope labeling by amino acids in cell culture (SILAC) of a human virus. Inactivation by UVC light and the full sunlight spectrum were found to efficiently inactivate HAdV, whereas UVA-visible light only caused inactivation in the presence of external sensitizers (indirect solar disinfection). Genome damage was significant for UVC but was less important for solar disinfection. In contrast, indirect solar disinfection exhibited extensive protein degradation. In particular, the fiber protein and the amino acids responsible for host binding within the fiber protein were shown to degrade. In addition, the central domain of the penton protein was damaged, which may inhibit interactions with the fiber protein and lead to a disruption of the initial stages of infection. Damage to the hexon protein, however, appeared to affect only regions not directly involved in the infectious cycle.