Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag.
Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag.
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DOI：
10.3390/v8090256
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发表时间：
2016-09-19
期刊：
影响因子：
--
通讯作者：
Musier-Forsyth K
中科院分区：
文献类型：
--
作者：
Rye-McCurdy T;Olson ED;Liu S;Binkley C;Reyes JP;Thompson BR;Flanagan JM;Parent LJ;Musier-Forsyth K
逆转录病毒特别包装全长的二聚体基因组RNA(GRNA)，即使在细胞RNA大量过剩的情况下也是如此。位于gRNA5‘-非翻译区(5’非翻译区)的“psi”(Ψ)元件通过与GAG的核衣壳(NC)结构域相互作用而对包装起关键作用。然而，在体外，Ψ和非ΨRNA的GAG结合亲和力没有显著差异。以往的盐滴定结合实验表明，人类免疫缺陷病毒1型Gag与Ψ核糖核酸结合具有较高的特异性和较少的电荷相互作用，而与非Ψ核糖核酸结合的特异性较低，涉及更多的静电作用。NC结构域对于特定的Ψ结合是至关重要的，但令人惊讶的是，缺少基质(MA)结构域的Gag突变体在区分Ψ和非ΨRNA时效率较低。我们现在发现，劳斯肉瘤病毒Gag也以MA依赖的方式有效地区分RSVRNA和非RNARNA。有趣的是，HIV-1和RSVMA结构域互换的Gag嵌合体与同源ΨRNA保持了高的结合特异性。利用Ψ核糖核酸突变结构，在两个系统中都确定了负责促进高GAG结合特异性的决定因素。综上所述，这些研究揭示了HIV-1和RSVMA结构域在促进GAG对ΨRNA的选择性方面的功能等价性，以及促进这种选择性的Ψ元件。
Retroviruses specifically package full-length, dimeric genomic RNA (gRNA) even in the presence of a vast excess of cellular RNA. The “psi” (Ψ) element within the 5′-untranslated region (5′UTR) of gRNA is critical for packaging through interaction with the nucleocapsid (NC) domain of Gag. However, in vitro Gag binding affinity for Ψ versus non-Ψ RNAs is not significantly different. Previous salt-titration binding assays revealed that human immunodeficiency virus type 1 (HIV-1) Gag bound to Ψ RNA with high specificity and relatively few charge interactions, whereas binding to non-Ψ RNA was less specific and involved more electrostatic interactions. The NC domain was critical for specific Ψ binding, but surprisingly, a Gag mutant lacking the matrix (MA) domain was less effective at discriminating Ψ from non-Ψ RNA. We now find that Rous sarcoma virus (RSV) Gag also effectively discriminates RSV Ψ from non-Ψ RNA in a MA-dependent manner. Interestingly, Gag chimeras, wherein the HIV-1 and RSV MA domains were swapped, maintained high binding specificity to cognate Ψ RNAs. Using Ψ RNA mutant constructs, determinants responsible for promoting high Gag binding specificity were identified in both systems. Taken together, these studies reveal the functional equivalence of HIV-1 and RSV MA domains in facilitating Ψ RNA selectivity by Gag, as well as Ψ elements that promote this selectivity.
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作者：
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发表时间：
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期刊：
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