确定RNA结构多样性及其在HIV-1RNA拼接中的作用
Phillip J Tomezsko1,2,3, Vincent D A Corbin4,5, Paromita Gupta1
1Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Nature
|June 20, 2020
概括
研究人类免疫缺陷病毒1 (HIV-1) 的RNA结构,发现结合部位的异质区域和替代形状. 这一发现支持RNA结构异质性调节病毒基因表达和拼接的假设.
科学领域:
- 分子生物学
- 病毒学
- 遗传学
背景情况:
- 人类免疫缺陷病毒1 (HIV-1) 依赖于其单链RNA基因组的替代拼接来产生多种蛋白质.
- 在HIV-1中控制替代拼接位选择的精确机制在很大程度上是未知的.
- 影响拼接和病毒复制的同名RNA突变表明存在未表征的cis-regulatory元素.
研究的目的:
- 研究HIV-1RNA基因组的细胞内结构.
- 识别替代RNA构造及其在调节结合位点选择中的作用.
- 为RNA结构异质性影响病毒基因表达的假设提供证据.
主要方法:
- 使用二甲基硫酸盐突变分析与测序 (DMS- MaPseq) 来探测细胞环境中的HIV-1RNA结构.
- 一种新的算法"使用预期最大化检测RNA折叠组合" (DREEM) 已被开发用于分析RNA结构异质性.
- 同时测量细胞内RNA结构和拼接模式.
主要成果:
- 分析显示了整个HIV-1基因组中的异质结构区域,挑战了以前的人口平均模型.
- 在关键拼接部位确定了替代RNA构造,影响了转录异型的比率.
- 这项研究证实了先前在HIV-1 Rev响应元素中特征化的细胞内部结构的存在.
结论:
- 细胞内HIV-1RNA存在于异构的结构中,而不是单一的平均结构.
- 在接合部位的替代RNA结构直接影响HIV-1替代接合的调节.
- 这项工作提供了强有力的证据,表明RNA构型异质性是HIV-1基因表达和拼接的关键调节者.
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