CHARACTERIZATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 DIMERIC RNA FROM WILD-TYPE AND PROTEASE-DEFECTIVE VIRIONS
CHARACTERIZATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 DIMERIC RNA FROM WILD-TYPE AND PROTEASE-DEFECTIVE VIRIONS
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DOI：
10.1128/jvi.68.8.5013-5018.1994
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发表时间：
1994-08-01
影响因子：
5.4
通讯作者：
REIN, A
中科院分区：
文献类型：
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作者：
FU, W;GORELICK, RJ;REIN, A
我们已经鉴定了野生型和缺乏蛋白酶(PR)的人类免疫缺陷病毒1型(HIV-1)颗粒中的二聚体基因组RNA。我们发现从PR突变型病毒粒子中分离的二聚体RNA在非变性凝胶电泳法中的迁移率低于野生型病毒粒子。它还在比野生型二聚体更低的温度下分解成单体。因此，PR(-)粒子中的二聚体与野生型粒子中的二聚体具有不同的构象。这些结果与最近对Moloney鼠白血病病毒的研究结果非常相似，表明组装后依赖PR的成熟事件是逆转录病毒基因组RNA的共同特征。我们还测量了野生型和PR(-)二聚体RNA在不同离子条件下的热稳定性。这两种形式的二聚体都通过增加Na+浓度而稳定下来。然而，孵化缓冲液中存在的单价阳离子的一致性对这两种形式的熔融温度没有显著影响。这一观察结果与最近关于由HIV-1序列的短片段在体外形成的二聚体的报道形成了对比：后者的二聚体特定地被K+离子稳定。K+在体外形成的二聚体的稳定性被认为是鸟嘌呤四元结构存在的证据。结果表明，鸟嘌呤四元组不参与将HIV-1全长、真实基因组RNA连接成二聚体结构的结构。
We have characterized the dimeric genomic RNA in particles of both wild-type and protease (PR)-deficient human immunodeficiency virus type 1 (HIV-1). We found that the dimeric RNA isolated from PR- mutant virions has a lower mobility in nondenaturing gel electrophoresis than that from wild-type virions. It also dissociates into monomers at a lower temperature than the wild-type dimer. Thus, the dimer in PR(-) particles is in a conformation different from that in wild-type particles. These results are quite similar to recent findings on Moloney murine leukemia virus and suggest that a postassembly, PR-dependent maturation event is a common feature in genomic RNAs of retroviruses. We also measured the thermal stability of the wild-type and PR(-) dimeric RNAs under different ionic conditions. Both forms of the dimer were stabilized by increasing Na+ concentrations. However, the melting temperatures of the two forms were not significantly affected by the identity of the monovalent cation present in the incubation buffer. This observation is in contrast with recent reports on dimers formed in vitro from short segments of HIV-1 sequence: the latter dimers are specifically stabilized by K+ ions. K+ stabilization of dimers formed in vitro has been taken as evidence for the presence of guanine quartet structures. The results suggest that guanine quartets are not involved in the structure linking full-length, authentic genomic RNA of HIV-1 into a dimeric structure.