VIRAL-RNA ANNEALING ACTIVITIES OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 NUCLEOCAPSID PROTEIN REQUIRE ONLY PEPTIDE DOMAINS OUTSIDE THE ZINC FINGERS
VIRAL-RNA ANNEALING ACTIVITIES OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 NUCLEOCAPSID PROTEIN REQUIRE ONLY PEPTIDE DOMAINS OUTSIDE THE ZINC FINGERS
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DOI：
10.1073/pnas.89.14.6472
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发表时间：
1992-07-15
影响因子：
11.1
通讯作者：
DARLIX, JL
中科院分区：
文献类型：
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作者：
DEROCQUIGNY, H;GABUS, C;DARLIX, JL
1型人类免疫缺陷病毒的核衣壳（NC）由大量NC蛋白分子组成，可能将二聚体RNA基因组包裹在病毒粒子内核内。 NC 蛋白是一种 gag 编码产物，包含两个侧翼为碱性残基的锌指。在人类免疫缺陷病毒 1 型病毒体中，NCp15 最终被加工成 NCp7 和 p6 蛋白。在病毒粒子组装过程中，逆转录病毒 NC 蛋白对于核心形成和基因组 RNA 衣壳化是必需的，这对于病毒感染性至关重要。体外 NCp15 激活病毒 RNA 二聚化，这一过程在体内最有可能与基因组 RNA 包装以及复制引物 tRNA(Lys,3) 退火至逆转录起始位点有关。为了表征人类免疫缺陷病毒 1 型 NC 蛋白的各种功能所需的结构域，以纯形式合成了 72 个氨基酸的 NCp7 和几种衍生肽。我们在此表明，具有或不具有两个锌指的合成 NCp7 具有 NCp15 的 RNA 退火活性。进一步删除 N 端 12 个氨基酸和 C 端 8 个氨基酸，导致缺乏指结构域的 27 个残基肽，对 NC 蛋白的体外活性影响很小或没有影响。然而，删除第一个振铃侧翼含有碱性残基的短序列会导致 NC 蛋白活性完全丧失。提出碱性残基和锌指配合在体内选择和包装基因组RNA。因此，与 NC 蛋白第一个锌指侧翼的碱性残基相关的病毒 RNA 结合和退火活性的抑制可用作抗病毒剂设计的模型。
The nucleocapsid (NC) of human immunodeficiency virus type 1 consists of a large number of NC protein molecules, probably wrapping the dimeric RNA genome within the virion inner core. NC protein is a gag-encoded product that contains two zinc fingers flanked by basic residues. In human immunodeficiency virus type 1 virions, NCp15 is ultimately processed into NCp7 and p6 proteins. During virion assembly the retroviral NC protein is necessary for core formation and genomic RNA encapsidation, which are essential for virus infectivity. In vitro NCp15 activates viral RNA dimerization, a process most probably linked in vivo to genomic RNA packaging, and replication primer tRNA(Lys,3) annealing to the initiation site of reverse transcription. To characterize the domains of human immunodeficiency virus type 1 NC protein necessary for its various functions, the 72-amino acid NCp7 and several derived peptides were synthesized in a pure form. We show here that synthetic NCp7 with or without the two zinc fingers has the RNA annealing activities of NCp15. Further deletions of the N-terminal 12 and C-terminal 8 amino acids, leading to a 27-residue peptide lacking the finger domains, have little or no effect on NC protein activity in vitro. However deletion of short sequences containing basic residues flanking the first ringer leads to a complete loss of NC protein activity. It is proposed that the basic residues and the zinc fingers cooperate to select and package the genomic RNA in vivo. Inhibition of the viral RNA binding and annealing activities associated with the basic residues flanking the first zinc finger of NC protein could therefore be used as a model for the design of antiviral agents.