State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China.
Int J Mol Sci. 2016 Feb 26;17(3):252. doi: 10.3390/ijms17030252.
Studies of the targets of anti-viral compounds are hot topics in the field of pesticide research. Various efficient anti-TMV (Tobacco Mosaic Virus) compounds, such as Ningnanmycin (NNM), Antofine (ATF), Dufulin (DFL) and Bingqingxiao (BQX) are available. However, the mechanisms of the action of these compounds on targets remain unclear. To further study the mechanism of the action of the anti-TMV inhibitors, the TMV coat protein (TMV CP) was expressed and self-assembled into four-layer aggregate disks in vitro, which could be reassembled into infectious virus particles with TMV RNA. The interactions between the anti-TMV compounds and the TMV CP disk were analyzed by size exclusion chromatography, isothermal titration calorimetry and native-polyacrylamide gel electrophoresis methods. The results revealed that assembly of the four-layer aggregate disk was inhibited by NNM; it changed the four-layer aggregate disk into trimers, and affected the regular assembly of TMV CP and TMV RNA. The four-layer aggregate disk of TMV CP was little inhibited by ATF, DFL and BQX. Our results provide original data, as well as new strategies and methods, for research on the mechanism of action of anti-viral drugs.
抗病毒化合物作用靶标的研究是农药领域的热点。已开发出各种高效的抗 TM V（烟草花叶病毒）化合物，如宁南霉素（NNM）、抗毒威（ATF）、毒氟磷（DFL）和病毒必克（BQX）。然而，这些化合物对靶标的作用机制尚不清楚。为了进一步研究抗病毒抑制剂的作用机制，本研究在体外表达了 TM V 外壳蛋白（TMV CP）并自组装成四层聚集体盘，该聚集体盘可与 TM V RNA 重新组装成感染性病毒颗粒。采用排阻色谱、等温滴定量热法和非变性聚丙烯酰胺凝胶电泳方法分析了抗病毒化合物与 TM V CP 盘之间的相互作用。结果表明，NNM 抑制了四层聚集体盘的组装；它将四层聚集体盘转变成三聚体，影响了 TM V CP 和 TM V RNA 的规则组装。ATF、DFL 和 BQX 对 TM V CP 的四层聚集体盘的抑制作用很小。本研究结果为抗病毒药物作用机制的研究提供了原始数据，以及新的策略和方法。
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