靶向G-四链体的抗番茄丛矮病毒卟啉类衍生物的筛选  

Screening of anti-tomato bushy stunt virus porphyrin derivatives targeting G-quadruplex

作　　者：谢从宝 祝强强 张涛[7] 宁丽红[7] 位灯国 XIE Congbao;ZHU Qiangqiang;ZHANG Tao;NING Lihong;WEI Dengguo(National Key Laboratory for the Exploitation and Utilization of Agricultural Microbial Resources,College of Vetrinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;Hubei Hongshan Laboratory,Huazhong Agricultural University,Wuhan 430070,China;National Reference Laboratory of Veterinary Drug Residues and MAO Key Laboratory for Detection of Veterinary Drug Residues,Wuhan 430070,China;Shenzhen Institute of Nutrition and Health,Huazhong Agricultural University,Shenzhen 518000,China;Shenzhen Branch,Guangdong Laboratory for Lingnan Modern Agriculture,Genome Analysis Laboratory of the Ministry of Agriculture,Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518000,China;Interdisciplinary Sciences Institute,Huazhong Agricultural University,Wuhan 430070,China;College of Science,Huazhong Agricultural University,Wuhan 430070,China;Key Laboratory of Horticulture Biology,Ministry of Education,College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学农业微生物资源发掘与利用全国重点实验室,动物医学院,武汉430070 [2]湖北洪山实验室,武汉430070 [3]华中农业大学,兽药残留国家基准实验室及农业部兽药残留检测重点实验室,武汉430070 [4]华中农业大学深圳营养与健康研究院,深圳518000 [5]中国农业科学院深圳农业基因组研究所和岭南现代农业科学与技术广东省实验室深圳分中心,深圳518000 [6]华中农业大学交叉科学研究院,武汉430070 [7]华中农业大学理学院,武汉430070 [8]华中农业大学园艺植物生物学教育部重点实验室,园艺林学学院,武汉430070

出　　处：《农药学学报》2023年第2期319-328,352,共11页Chinese Journal of Pesticide Science

基　　金：国家自然科学基金(21732002,22077043,31672558);华中农业大学-中国农业科学院深圳农业基因组研究所合作基金(SZYJY2022016);湖北省杰出青年基金(2021CFA061).

摘　　要：植物病毒病给蔬菜生产带来很大影响,目前蔬菜病毒防控主要采用非药剂措施,广谱抗病毒剂效果有限,需要基于新的药物靶点开发新的病毒抑制剂。G-四链体(G4s)是一种特殊的核酸高级结构,其在病毒基因组中的形成或解链可调节基因的复制、转录和翻译等过程,进而影响病毒增殖。一些可调节G-四链体结构稳定性的小分子呈现出抗病毒活性,使得G-四链体有望成为新的抗病毒药物的靶标。番茄丛矮病毒(tomato bushy stunt virus,TBSV)是一种分布广泛的植物病毒,有关其基因组的复制、转录和翻译等分子机制已研究得较为成熟,是一种研究病毒与寄主互作的模式病毒。揭示TBSV中G-四链体的结构及功能,有望为植物病毒基因中G-四链体的研究奠定基础。本研究通过生物信息学分析,在TBSV基因中鉴定出两条保守的、潜在的G-四链体可形成序列(putative G-quadruplex sequences,PQS)——TBSV-PQS2和TBSV-PQS4;通过紫外、荧光光谱和圆二色光谱(CD光谱)方法筛选出与TBSV-PQS2和TBSV-PQS4互作的G4配体;通过烟草体内侵染性克隆试验发现,G-四链体配体N-甲基中卟啉IX(NMM)可抑制TBSV病毒的增殖。本研究结果表明,以植物病毒中G-四链体为靶点开发抗植物病毒的生态友好型农药,有望成为控制植物病毒危害的新策略。Plant viruses have a great adverse impact on vegetable production.Non-pharmaceutical measures are mainly adopted for vegetable virus prevention and control at present.However,the efficacy of broad-spectrum antiviral agents is limited.The new viral inhibitors are expected to be developed based on novel drug targets.G-quadruplex(G4s)is a non-canonical nucleic acid structure.The replication,transcription,and translation of genes can be regulated by the formation or unfolding of G-quadruplexes in the genome,which affects virus proliferation.Small molecules mediating the stability of G-quadruplex structures exhibit antiviral activity,making G-quadruplexes to be potential antiviral targets.As a widely distributed plant virus,tomato bushy stunt virus(TBSV)is considered as a model virus to study the interaction between the virus and host.The molecular mechanisms of its genome replication,transcription,and translation have been widely studied.Revealing the structure and function of G-quadruplex in TBSV is expected to lay a foundation for the research of G-quadruplex in plant virus genes.In this work,two conserved potential G-quadruplex sequences(putative G-quadruplex sequences,PQS)TBSV-PQS2 and TBSV-PQS4 were identified in TBSV genes through bioinformatics analysis.The G4 ligands interacting with TBSV-PQS2 and TBSV-PQS4 were screened out by UV fluorescence spectrum and circular dichroism(CD)spectrum.Invasive cloning experiments in tobacco found that G-quadruplex ligand N-methyl mesophorphyrin IX(NMM)inhibits TBSV virus proliferation.This work shows that the development of ecological pesticides against plant viruses by targeting the G-quadruplex in plant viruses is expected to become a new strategy for controlling the damage of plant viruses.

关 键 词：RNA G-四链体 植物病毒 番茄丛矮病毒(TBSV) N-甲基中卟啉IX(NMM) 

分 类 号：TQ450.1[化学工程—农药化工]