Structural Organizations of Qβ and MS2 Phages Affect Capsid Protein Modifications by Oxidants Hypochlorous Acid and Peroxynitrite
Structural Organizations of Qβ and MS2 Phages Affect Capsid Protein Modifications by Oxidants Hypochlorous Acid and Peroxynitrite
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DOI：
10.3389/fmicb.2020.01157
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发表时间：
2020-06-03
影响因子：
5.2
通讯作者：
Gantzer, Christophe
中科院分区：
文献类型：
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作者：
Bastin, Guillaume;Loison, Pauline;Gantzer, Christophe
致病性肠道病毒和噬菌体如Q β和MS2通过粪-口途径传播。然而，氧化剂如过氧亚硝酸盐（ONOOH）和次氯酸（HClO）可以通过灭活传染性病毒来预防新的感染。它们的杀病毒作用是公认的，但预测氧化剂对病毒的作用目前是不可能的，因为病毒灭活的详细机制尚不清楚。我们的数据显示ONOOH和HClO交联了Q β和MS2噬菌体的衣壳蛋白和RNA基因组。在透射电子显微镜（TEM）下，即使99%的噬菌体被氧化灭活，衣壳仍然完好无损。此外，对衣壳蛋白的精确分子研究表明，ONOOH和HClO优先靶向含有氧化敏感氨基酸C、Y或w的衣壳蛋白区域。有趣的是，这些氨基酸的相互作用是决定它们是否会被O、Cl或NO(2)修饰，或者是否会导致质谱检测到的蛋白质区域的损失的关键参数，它们共同提示了交联形成的潜在位点。总之，这些数据表明，HClO和ONOOH一致地靶向氧化敏感氨基酸，而不考虑Q β和MS2的结构组织，即使表型随着与邻近蛋白质/RNA的相互作用而改变。这些数据还表明了一种潜在的病毒失活的新机制，其中交联可能损害传染性。
Pathogenic enteric viruses and bacteriophages such as Q beta and MS2 are transmitted through the fecal-oral route. However, oxidants such as peroxynitrite (ONOOH) and hypochlorous acid (HClO) can prevent new infection by inactivating infectious viruses. Their virucidal effect is well recognized, and yet predicting the effects of oxidants on viruses is currently impossible because the detailed mechanisms of viral inactivation remain unclear. Our data show that ONOOH and HClO cross-linked the capsid proteins and RNA genomes of Q beta and MS2 phages. Consistently, the capsids appeared intact by transmission electron microscopy (TEM) even when 99% of the phages were inactivated by oxidation. Moreover, a precise molecular study of the capsid proteins shows that ONOOH and HClO preferentially targeted capsid protein regions containing the oxidant-sensitive amino acid C, Y, or W. Interestingly, the interaction of these amino acids was a crucial parameter defining whether they would be modified by the addition of O, Cl, or NO(2)or whether it induced the loss of the protein region detected by mass spectrometry, together suggesting potential sites for cross-link formation. Together, these data show that HClO and ONOOH consistently target oxidant-sensitive amino acids regardless of the structural organization of Q beta and MS2, even though the phenotypes change as a function of the interaction with adjacent proteins/RNA. These data also indicate a potential novel mechanism of viral inactivation in which cross-linking may impair infectivity.