Mutational analysis of two zinc finger motifs in HIV type 1 nucleocapsid proteins: effects on proteolytic processing of Gag precursors and particle formation.
Mutational analysis of two zinc finger motifs in HIV type 1 nucleocapsid proteins: effects on proteolytic processing of Gag precursors and particle formation.
复制标题
HIV 1 型核衣壳蛋白中两个锌指基序的突变分析：对 Gag 前体蛋白水解加工和颗粒形成的影响。
DOI：
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发表时间：
1996
影响因子：
1.5
通讯作者：
Masanori Hayami
中科院分区：
文献类型：
--
作者：
Akiko Mizuno;Eiji Ido;Toshiyuki Goto;T. Kuwata;M. Nakai;Masanori Hayami
为了阐明人类免疫缺陷病毒1型（HIV-1） Gag蛋白核衣壳（NC）结构域的两个锌指基序的生理功能，我们通过定点诱变将两个基序中的任一基序中的半胱氨酸变为丝氨酸或将两者都变为丝氨酸。任何一种突变都失去了病毒的传染性，但它们的影响在各自的突变体中表现不同。Northern blot分析表明，第一个手指突变体在基因组RNA封装方面的效率要低得多（约为野生型的10%），而两个手指的双突变体完全无法封装RNA。相比之下，第二指突变体保留了其RNA封装能力，其效率与野生型相似。转染突变前病毒dna的cd4阳性M8166细胞裂解物的免疫印迹分析显示，在第一指序列发生改变的两种突变病毒中，Gag前体的加工被延迟，而第二指突变病毒的加工似乎是正常的。另一方面，对病毒颗粒的免疫印迹分析显示，第二指突变颗粒含有一些被认为是p24CA降解产物的蛋白质。电镜观察表明，这些突变病毒的颗粒除了直径略大于野生型外，在形态上都是相似的。这些结果表明HIV-1 NC蛋白的这些手指基序的功能并不相同。也就是说，第一根手指主要负责RNA的封装，第二根手指则是稳定病毒颗粒所必需的。
To clarify the physiological function of two zinc finger motifs in the nucleocapsid (NC) domain of the Gag protein of human immunodeficiency virus type 1 (HIV-1), we changed cysteine to serine in either of the two motifs or both by site-directed mutagenesis. Viral infectivity was lost by any of the mutations, but their effects appeared differently in the respective mutants. Northern blot analysis showed that the first finger mutant was far less efficient (approximately 10% of the wild type) in genomic RNA encapsidation and that the dual mutant of both fingers completely failed to encapsidate the RNA. In contrast, the second finger mutant retained its ability for RNA encapsidation with an efficiency similar to that of the wild type. Immunoblot analysis of the lysates of CD4-positive M8166 cells transfected with the mutant proviral DNAs showed that the processing of Gag precursors was delayed in two mutant viruses having alterations in the first finger sequence, whereas the processing of the second finger mutant appeared to be normal. On the other hand, immunoblot analysis of the virus particles showed that the second finger mutant particles contained some proteins that were thought to be degradation products of p24CA. Electron microscopic observation showed that all particles of these mutant viruses were morphologically alike except that they had a slightly larger diameter than that of the wild type. These results indicate that these finger motifs of HIV-1 NC protein do not function equivalently. Namely, the first finger is primarily responsible for RNA encapsidation and the second is required for stabilization of virus particles.
DOI：
10.1073/pnas.85.22.8420
发表时间：
1988-11-01
影响因子：
11.1
作者：
GORELICK, RJ;HENDERSON, LE;REIN, A
通讯作者：
REIN, A
禽逆转录病毒核衣壳蛋白的定点诱变，第 12 页。影响体外 RNA 结合的突变会阻止病毒复制。
DOI：
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发表时间：
1988
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Fu,XD;Katz,RA;Skalka,AM;Leis,J
通讯作者：
Leis,J