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对猫免疫缺陷病毒基因组RNA包装至关重要的假定Gag结合部位的识别

Anjana Krishnan¹, Lizna M Ali¹, Suresha G Prabhu¹

¹Department of Microbiology and Immunology, College of Medicine and Health Sciences (CMHS), United Arab Emirates University (UAEU), Al Ain, United Arab Emirates.

RNA (New York, N.Y.)

|November 1, 2023

概括

猫类免疫缺陷病毒 (FIV) 包装信号 (ψ) 需要一个特定的U$^{85}$CUG$^{88}$序列来有效地包装基因组RNA (gRNA). 这一序列对于Gag蛋白结合和gRNA封装到新病毒中至关重要.

科学领域:

病毒学病毒学
分子生物学分子生物学
结构生物学结构生物学

背景情况:

逆转录病毒Gag蛋白通过结合包装信号 (ψ) 来选择和包装病毒基因组RNA (gRNA).
猫类免疫缺陷病毒 (FIV) 包装信号 (ψ) 位于gRNA的5'端,形成一个复杂的高级结构.
之前的研究将FIV ψ映射到两个不连续的区域,需要对关键结构元素进行进一步调查.

研究的目的:

精确定义FIV包装信号 (ψ) 区域和gRNA包装必不可少的结构动机.
阐明FIV ψ内特定序列在gRNA封装和Gag蛋白相互作用中的作用.
探索已识别的序列作为逆转录病毒疗法和基于FIV的基因治疗载体的标的潜力.

主要方法:

对FIV ψ序列的系统突变分析,以评估它们对gRNA包装的影响.
高通量选择性2'酸乙化通过原料扩展 (hSHAPE) 分析,以评估野生类型和突变型 ψ RNA 的结构变化.
使用纯化的FIV Pr50^Gag进行过器结合试验,以确定与野生类型和突变型 ψ RNA 的结合亲和力.

主要成果:

突变分析发现,在FIV ψ内,一个未配对的U$^{85}$CUG$^{88延伸对于gRNA封装至关重要.
hSHAPE测试表明,U$^{85}$CUG$^{88的突变并没有显著改变整体RNA结构,保持80-92核酸不配对.
波器结合试验表明,FIV Pr50^Gag与突变U$^{85}$CUG$^{88 ψRNAs的结合减少,证实了该序列对Gag-ψRNA相互作用的重要性.

结论:

单链U$^{85}$CUG$^{88序列对于FIV gRNA包装至关重要,主要介导与Gag蛋白的相互作用.
尽管有影响包装的突变,但 ψ RNA 的结构完整性仍然保持不变,突出显示了 U$^{85}$CUG$^{88}$ 的序列特定作用.
了解这些FIV gRNA包装决定因素可以促进开发新的抗病毒策略和基于FIV的基因治疗载体.

关键词:

口/RNA相互作用 RNA包装信号 (ψ) 是一个RNA包装信号.RNA的二级结构是RNA的二级结构.猫类免疫缺陷病毒病毒.过器结合测定试验这是一个hSHAPE.它们是逆转录病毒.

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