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首页> 外文期刊>Journal of Molecular Biology >Display of epitopes on the surface of tobacco mosaic virus: impact of charge and isoelectric point of the epitope on virus-host interactions.
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Display of epitopes on the surface of tobacco mosaic virus: impact of charge and isoelectric point of the epitope on virus-host interactions.

机译：烟草花叶病毒表面上表位的展示：表位的电荷和等电点对病毒-宿主相互作用的影响。

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摘要

The biophysical properties of the tobacco mosaic tobamovirus (TMV) coat protein (CP) make it possible to display foreign peptides on the surface of TMV. The immunogenic epitopes G5-24 from the rabies virus (RV) glycoprotein, and 5B19 from murine hepatitis virus (MHV) S-glycoprotein were successfully displayed on the surface of TMV, and viruses accumulated to high levels in infected leaves of Nicotiana tabacum Xanthi-nn. The peptide RB19, which contains an arginine residue plus the 5B19 epitope fused to the CP (TMV-RB19), resulted in the induction of necrotic local lesions on inoculated leaves of N. tabacum Xanthi-nn and cell death of infected BY2 protoplasts. RNA dot blot assays confirmed that expression of the acidic and basic pathogenesis-related PR2 genes were induced in infected Xanthi-nn leaf tissue. TMV that carried epitope 31D from the RV nucleoprotein did not accumulate in inoculated tobacco leaves. Analysis of hybrid CPs predicted that the isoelectric points (pI):charge value was 5.31:-2 for wild-type CP, 5.64:-1 for CP-RB19, and 9.14:+2 for CP-31D. When acidic amino acids were inserted in CP-RB19 and CP-31D to bring their pI:charge to near that of wild-type CP, the resulting viruses TMV-RB19E and TMV-4D:31D infected N. tabacum Xanthi-nn plants and BY2 protoplasts without causing cell death. These data show the importance of the pI of the epitope and its effects on the hybrid CP pI:charge value for successful epitope display as well as the lack of tolerance to positively charged epitopes on the surface of TMV. Copyright 1999 Academic Press.
机译：烟草花叶病毒Tobamovirus（TMV）外壳蛋白（CP）的生物物理特性使其可以在TMV表面展示外源肽。狂犬病毒（RV）糖蛋白的免疫原性表位G5-24和鼠肝炎病毒（MHV）S-糖蛋白的5B19成功展示在TMV的表面上，并且病毒在烟草Xanthi- nn。含有精氨酸残基和与CP融合的5B19表位的肽RB19（TMV-RB19）导致了烟草Xanthi-nn接种叶片上坏死局部损伤的诱导和被感染的BY2原生质体的细胞死亡。 RNA点印迹分析证实了在感染的Xanthi-nn叶组织中诱导了酸性和基本的发病机理相关PR2基因的表达。带有RV核蛋白表位31D的TMV不会在接种的烟叶中积累。混合CP的分析预测，对于野生型CP，等电点（pI）：电荷值为5.31：-2，对于CP-RB19为5.64：-1，对于CP-31D为9.14：+2。当将酸性氨基酸插入CP-RB19和CP-31D中以使其pI：电荷接近野生型CP时，产生的病毒TMV-RB19E和TMV-4D：31D感染了烟草Xanthi-nn植物和BY2原生质体而不会引起细胞死亡。这些数据显示了表位pI的重要性及其对杂种CP pI：电荷值的影响，以成功显示表位，以及对TMV表面带正电荷的表位缺乏耐受性。版权所有1999，学术出版社。

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