Changes in the Genome of the Tick-Borne Encephalitis Virus during Cultivation

IF 1.5 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biology Pub Date : 2024-04-09 DOI:10.1134/s0026893324020146
V. A. Ternovoi, E. P. Ponomareva, E. V. Protopopova, N. L. Tupota, T. P. Mikryukova, V. B. Loktev
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Abstract

The tick-borne encephalitis virus (TBEV) strain C11-13 (GenBank acc. no. OQ565596) of the Siberian genotype was previously isolated from the brain of a deceased person. TBEV C11-13 variants obtained at passages 3 and 8 in SPEV cells were inoculated into the brains of white mice for subsequent passages. Full genome sequences of all virus variants were analyzed by high-throughput sequencing. A total of 41 single nucleotide substitutions were found to occur mainly in the genes for the nonstructural proteins NS3 and NS5 (GenBank MF043953, OP902894, and OP902895), and 12 amino acid substitutions were identified in the deduced protein sequences. Reverse nucleotide and amino acid substitutions were detected after three passages through mouse brains. The substitutions restored the primary structures that were characteristic of the isolate C11-13 from a human patient and changed during the eight subsequent passages in SPEV cells. In addition, the 3′-untranslated region (3′-UTR) of the viral genome increased by 306 nt. The Y3 and Y2 3'-UTR elements were found to contain imperfect L and R repeats, which were probably associated with inhibition of cellular XRN1 RNase and thus involved in the formation of subgenomic flaviviral RNAs (sfRNAs). All TBEV variants showed high-level reproduction in both cell cultures and mouse brains. The genomic changes that occurred during successive passages of TBEV are most likely due to its significant genetic variability, which ensures its efficient reproduction in various hosts and its broad distribution in various climatic zones.

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蜱传脑炎病毒基因组在培养过程中的变化
摘要 先前从一名死者的脑中分离出了西伯利亚基因型的蜱传脑炎病毒(TBEV)C11-13株(GenBank编号:OQ565596)。将在 SPEV 细胞中第 3 和第 8 个传代获得的 TBEV C11-13 变体接种到白鼠脑中进行后续传代。通过高通量测序分析了所有病毒变体的全基因组序列。共发现 41 个单核苷酸置换,主要发生在非结构蛋白 NS3 和 NS5 的基因中(GenBank MF043953、OP902894 和 OP902895),在推导出的蛋白质序列中发现 12 个氨基酸置换。在小鼠大脑中进行三次传递后,检测到了反向核苷酸和氨基酸置换。这些置换恢复了来自人类患者的 C11-13 分离物的主要结构特征,并在随后的八次 SPEV 细胞传代中发生了变化。此外,病毒基因组的 3′-非翻译区(3′-UTR)增加了 306 nt。研究发现,Y3和Y2 3'-UTR元件含有不完全的L和R重复序列,这可能与抑制细胞XRN1 RNase有关,从而参与了亚基因组黄病毒RNA(sfRNA)的形成。所有 TBEV 变体在细胞培养物和小鼠大脑中都表现出高水平的繁殖能力。TBEV在连续传代过程中发生的基因组变化很可能是由于其显著的遗传变异性,这确保了其在不同宿主体内的高效繁殖以及在不同气候带的广泛分布。
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来源期刊
Molecular Biology
Molecular Biology 生物-生化与分子生物学
CiteScore
1.30
自引率
8.30%
发文量
78
审稿时长
3 months
期刊介绍: Molecular Biology is an international peer reviewed journal that covers a wide scope of problems in molecular, cell and computational biology including genomics, proteomics, bioinformatics, molecular virology and immunology, molecular development biology, molecular evolution and related areals. Molecular Biology publishes reviews, experimental and theoretical works. Every year, the journal publishes special issues devoted to most rapidly developing branches of physical-chemical biology and to the most outstanding scientists.
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