Investigation of HIV-1 Gag binding with RNAs and lipids using Atomic Force Microscopy
Investigation of HIV-1 Gag binding with RNAs and lipids using Atomic Force Microscopy
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DOI：
10.1371/journal.pone.0228036
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发表时间：
2020-02-03
期刊：
影响因子：
3.7
通讯作者：
Mohideen, Umar
中科院分区：
文献类型：
--
作者：
Chen, Shaolong;Xu, Jun;Mohideen, Umar
利用原子力显微镜在垂直分辨率为0.1A、横向分辨率为1nm的溶液环境中研究Gag、psi RNA及其与脂质结合复合物的形态。TARpolyA RNA作为RNA对照。所使用的脂质为磷酸-酰肌醇-(4,5)-二磷酸（PI(4,5)P2）。研究了特异复合物Gag-psi RNA、Gag-TARpolyA RNA、Gag-PI(4,5)P2和PI(4,5)P2-psi RNA- gag的形态。它们在带正电荷或负电荷的云母衬底上成像，这取决于所携带的净电荷。凝胶电泳证实，Gag及其配合物由单体、二聚体和四聚体组成。将特定的psi RNA添加到Gag中可以增加Gag的多聚性。发现非特异性TARpolyA RNA不会导致Gag多聚化的增加。在Gag中加入PI(4,5)P2可以增加Gag的多聚性，但其程度低于psi RNA。当psi RNA和PI(4,5)P2同时存在时，Gag会发生构象变化和更高程度的多聚。
Atomic Force Microscopy was utilized to study the morphology of Gag, psi RNA, and their binding complexes with lipids in a solution environment with 0.1A vertical and 1nm lateral resolution. TARpolyA RNA was used as a RNA control. The lipid used was phospha-tidylinositol-(4,5)-bisphosphate (PI(4,5)P2). The morphology of specific complexes Gag-psi RNA, Gag-TARpolyA RNA, Gag-PI(4,5)P2 and PI(4,5)P2-psi RNA-Gag were studied. They were imaged on either positively or negatively charged mica substrates depending on the net charges carried. Gag and its complexes consist of monomers, dimers and tetramers, which was confirmed by gel electrophoresis. The addition of specific psi RNA to Gag is found to increase Gag multimerization. Non-specific TARpolyA RNA was found not to lead to an increase in Gag multimerization. The addition PI(4,5)P2 to Gag increases Gag multimerization, but to a lesser extent than psi RNA. When both psi RNA and PI(4,5)P2 are present Gag undergoes comformational changes and an even higher degree of multimerization.