Analysis of hepatitis C virus core protein interaction domains.
Analysis of hepatitis C virus core protein interaction domains.
复制标题
丙型肝炎病毒核心蛋白相互作用域分析。
作者：
Olga Nolandt;Viktoria Kern;Hubert Mu;ller;E. Pfaff;L. Theilmann;R. Welker;Hans;usslich
丙型肝炎病毒（HCV）核心蛋白形成病毒的内部外壳，其包裹基因组RNA，并被包裹在宿主细胞来源的脂质膜中。作为单个衣壳蛋白，核心应该能够多聚化，但在表达HCV结构蛋白后产生病毒样颗粒的尝试尚未成功。本研究利用酵母双杂交系统分析了全长和截短的HCV核心蛋白的相互作用能力。全长核心含有或缺乏易位信号的E1糖蛋白不与全长或截短的核心蛋白相互作用。截短到N-末端122个氨基酸揭示了一个相互作用结构域，该结构域被定位到从氨基酸82-102的富含色氨酸的序列，并被称为主要的同型相互作用结构域。核心蛋白C端疏水片段（aa 122-172）不能与自身相互作用，但与主要的同型相互作用结构域反式相互作用（弱异型相互作用结构域）。当与GAL 4的DNA结合结构域融合时，在其N和C末端截短的核心蛋白（aa 46-102）具有反式激活作用。基于我们的研究结果，我们认为，C-末端段可能在顺式与主要的同型相互作用域，从而防止多聚化。还观察到与截短的固定化核心102结合的体外翻译蛋白质的核心-核心相互作用。然而，在这个系统中的相互作用是不太具体的，这表明蛋白质相互作用和可能的构象改变的核心可能是依赖于实验系统。
Hepatitis C virus (HCV) core protein forms the internal viral coat that encapsidates the genomic RNA and is enveloped in a host cell-derived lipid membrane. As the single capsid protein, core should be capable of multimerization but attempts to produce virus-like particles following expression of HCV structural proteins have not been successful. In this study, we have analysed the interaction capacity of full-length and truncated HCV core using the yeast two-hybrid system. Full-length core containing or lacking the translocation signal for the E1 glycoprotein did not interact with full-length or truncated core proteins. Truncation to the N-terminal 122 aa revealed an interaction domain which was mapped to the tryptophan-rich sequence from aa 82-102 and was termed the main homotypic interaction domain. The C-terminal hydrophobic fragment of core (aa 122-172) was incapable of interacting with itself but interacted with the main homotypic interaction domain in trans (the weak heterotypic interaction domain). Core proteins truncated at their N and C termini (aa 46-102) were trans-activating when fused to the DNA-binding domain of GAL4. Based on our results, we suggest that the C-terminal segment may interact in cis with the main homotypic interaction domain and thereby prevent multimerization. Core-core interaction was also observed for in vitro-translated proteins bound to truncated immobilized core 102. However, interaction was less specific in this system suggesting that protein interaction and possibly conformational alteration of core may be dependent on the experimental system.
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影响因子：
3.7
作者：
Moradpour, D;Englert, C;Wands, JR
通讯作者：
Wands, JR
影响因子：
10.5
作者：
DURFEE, T;BECHERER, K;ELLEDGE, SJ
通讯作者：
ELLEDGE, SJ
DOI：
10.1073/pnas.88.21.9578
发表时间：
1991-11-01
影响因子：
11.1
作者：
CHIEN, CT;BARTEL, PL;FIELDS, S
通讯作者：
FIELDS, S
分析昆虫细胞中表达的丙型肝炎病毒衣壳、E1 和 E2/NS1 蛋白。
影响因子：
3.7
作者：
Lanford,RE;Notvall,L;Chavez,D;White,R;Frenzel,G;Simonsen,C;Kim,J
通讯作者：
Kim,J