The Zinc Content of HIV-1 NCp7 Affects Its Selectivity for Packaging Signal and Affinity for Stem-Loop 3.
The Zinc Content of HIV-1 NCp7 Affects Its Selectivity for Packaging Signal and Affinity for Stem-Loop 3.
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HIV-1 NCp7 的锌含量影响其对包装信号的选择性和对茎环 3 的亲和力。
DOI：
10.3390/v13101922
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发表时间：
2021-09-24
期刊：
影响因子：
--
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Wang Y;Guo C;Wang X;Xu L;Li R;Wang J
人类免疫缺陷病毒(HIV)核衣壳蛋白(NC)是一种小的、高度碱性的蛋白，含有两个CCHC锌指基序，是在病毒成熟过程中从Gag多聚蛋白的NC结构域中切割出来的。我们先前报道，在大肠杆菌中过表达的重组HIV-1 Gag和NCp7分别含有两个和一个锌离子，并且Gag对包装信号(Psi)的选择性和对Psi的茎环(SL)-3的亲和力远远高于NCp7。在本研究中，我们制备了含有0(0NCp7)、1(NCp7)和2(2NCp7)锌离子的NCp7，并比较了它们的二级结构、Psi选择性和SL3亲和力。随着锌含量的降低，样品的有序化程度降低。与NCp7相比，2NCp7表现出更高的Psi选择性和SL3亲和力，与Gag相似，而0NCp7表现出更低的Psi选择性和SL3亲和力，与NCp7的H23和H44K双突变体相似，这表明Gag NC结构域和成熟NCp7的不同RNA结合特性可能至少部分是由于它们的锌含量不同造成的。本研究将有助于阐明锌在病毒生命周期中的关键作用，并有助于进一步研究从GAG的NC结构域到成熟的NCp7的功能转换。
The nucleocapsid (NC) protein of human immunodeficiency (HIV) is a small, highly basic protein containing two CCHC zinc-finger motifs, which is cleaved from the NC domain of the Gag polyprotein during virus maturation. We previously reported that recombinant HIV-1 Gag and NCp7 overexpressed in an E. coli host contains two and one zinc ions, respectively, and Gag exhibited much higher selectivity for packaging signal (Psi) and affinity for the stem-loop (SL)-3 of Psi than NCp7. In this study, we prepared NCp7 containing 0 (0NCp7), 1 (NCp7) or 2 (2NCp7) zinc ions, and compared their secondary structure, Psi-selectivity and SL3-affinity. Along with the decrease of the zinc content, less ordered conformations were detected. Compared to NCp7, 2NCp7 exhibited a much higher Psi-selectivity and SL3-affinity, similar to Gag, whereas 0NCp7 exhibited a lower Psi-selectivity and SL3-affinity, similar to the H23&H44K double mutant of NCp7, indicating that the different RNA-binding property of Gag NC domain and the mature NCp7 may be resulted, at least partially, from their different zinc content. This study will be helpful to elucidate the critical roles that zinc played in the viral life cycle, and benefit further investigations of the functional switch from the NC domain of Gag to the mature NCp7.
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DOI：
10.1002/anie.201713172
发表时间：
2018-03-01
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Deshmukh L;Tugarinov V;Appella DH;Clore GM
通讯作者：
Clore GM
影响因子：
4.8
作者：
Fenstermacher, Katherine J.;DeStefano, Jeffrey J.
通讯作者：
DeStefano, Jeffrey J.
影响因子：
4.2
作者：
Baum, MK;Shor-Posner, G;Campa, A
通讯作者：
Campa, A
HIV-1 Gag 和 NCp7 对包装信号的选择性、对茎环 3 的亲和力以及 Zn2 含量的比较。
影响因子：
3.9
作者：
Guo, Chao;Yao, Xiaohong;Wang, Ying
通讯作者：
Wang, Ying
影响因子：
3.5
作者：
OMICHINSKI, JG;CLORE, GM;GRONENBORN, AM
通讯作者：
GRONENBORN, AM