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运用计算和实验方法预测马铃薯卷叶病毒RNA依赖RNA聚合酶的单核苷酸多态性

Prediction of single nucleotide polymorphisms of RNA dependent RNA polymerase for the potato leafroll virus using computational and experimental approaches.

作者信息

Aseel Dalia G, Elmaghraby Ayaat M, El-Far Ali

机构信息

Plant Protection and Biomolecular Diagnosis Department, City of Scientific Research and Technological Applications (SRTA, City), Arid Lands Cultivation Research Institute (ALCRI),, New Borg El-Arab, 21934, Egypt.

Nucleic Acid Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications (SRTA, City), Alexandria, Egypt.

出版信息

Sci Rep. 2025 Aug 17;15(1):30121. doi: 10.1038/s41598-025-14436-8.

DOI:10.1038/s41598-025-14436-8
PMID:40820237
Abstract

Potato leafroll disease, caused by potato leafroll virus (PLRV), is one of the most devastating diseases, resulting in significant losses to the potato crop. Research on comprehensive studies examining the prediction of mutations in PLRV genes for this viral disease is limited. This study aims to quantify and characterize the accumulation of these genetic mutations by predicting SNPs for PLRV RNA-dependent RNA polymerase (RdRP) using computational approaches. We utilized advanced online protein prediction tools to explore the impact of DNA mutations on protein function and structure. These tools enabled us to assess the potential level and location of disorder within proteins, providing crucial insights into their behavior. In our study, the polymerase chain reaction (PCR) yielded a product corresponding to the PLRV-RdRp gene, measuring 450 base pairs (bp). This sequence has been documented in GenBank under the access number MT576073 and is cataloged in the UniProt database with the ID A0A8E6I3S8. The phylogenetic analysis of the PLRV-RdRp sequence involved a comparative study with reference sequences in the NCBI database. We focused on two notable predicted inversion mutations located at positions 93 (A93T) and 117 (K117G). Remarkably, the (Lys/Gly) mutation at position 117 exhibited a substantial disorder percentage of 61%, significantly higher than the reference sequence's 13.4%. These single nucleotide polymorphisms (SNPs) are not just statistical anomalies; they have real implications. They alter RdRp protein stability and physicochemical properties, altering pocket residues and influencing the protein's interaction with natural compounds. In conclusion, our findings provide a solid foundation for future research and development efforts focused at the rapid diagnosis and sustainable management of PLRV.

摘要

由马铃薯卷叶病毒(PLRV)引起的马铃薯卷叶病是最具毁灭性的病害之一,会给马铃薯作物造成重大损失。针对这种病毒性病害,关于全面研究PLRV基因突变预测的研究有限。本研究旨在通过使用计算方法预测PLRV RNA依赖的RNA聚合酶(RdRP)的单核苷酸多态性(SNP),来量化和表征这些基因突变的积累情况。我们利用先进的在线蛋白质预测工具来探索DNA突变对蛋白质功能和结构的影响。这些工具使我们能够评估蛋白质内无序的潜在水平和位置,为其行为提供关键见解。在我们的研究中,聚合酶链反应(PCR)产生了一个与PLRV-RdRp基因对应的产物,长度为450个碱基对(bp)。该序列已在GenBank中以登录号MT576073记录,并在UniProt数据库中以ID A0A8E6I3S8编目。PLRV-RdRp序列的系统发育分析涉及与NCBI数据库中的参考序列进行比较研究。我们重点关注位于第93位(A93T)和第117位(K117G)的两个显著预测倒位突变。值得注意的是,第117位的(赖氨酸/甘氨酸)突变表现出61%的显著无序百分比,明显高于参考序列的13.4%。这些单核苷酸多态性(SNP)并非只是统计异常;它们具有实际影响。它们改变了RdRp蛋白的稳定性和理化性质,改变了口袋残基并影响了该蛋白与天然化合物的相互作用。总之,我们的研究结果为未来致力于PLRV快速诊断和可持续管理的研发工作提供了坚实基础。

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