A large extension to HIV-1 Gag, like Pol, has negative impacts on virion assembly.
A large extension to HIV-1 Gag, like Pol, has negative impacts on virion assembly.
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DOI：
10.1371/journal.pone.0047828
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Morikawa Y
中科院分区：
文献类型：
--
作者：
Haraguchi H;Noda T;Kawaoka Y;Morikawa Y
HIV-1的GagPol蛋白含有病毒酶，如蛋白酶（PR）、逆转录酶和整合酶，这些酶对病毒粒子的感染性都至关重要。先前的研究表明，单独表达GagPol不会产生病毒颗粒，并且出芽缺陷是由Pol区域的存在引起的。然而，为什么GagPol不能产生病毒颗粒仍然是未知的。我们在这里表明，HIV-1 GagPol是无法膜结合和随后的颗粒组装。我们的共聚焦数据表明，尽管完全N-豆蔻酰化，GagPol蛋白未能靶向质膜与细胞质中的扩散分布。膜浮选分析证实了这些发现。GagPol的进行性C末端截短以得到GagPR允许质膜靶向，但仍然不允许颗粒产生。相反，C-末端添加的非同源蛋白质，如β-半乳糖苷酶或4串联GFP，Gag损害的膜亲和力，表明Pol区域，一个大的延伸Gag，抑制膜结合的背景下，GagPol。添加Fyn激酶[Fyn（10）]的10个N-末端氨基酸（一种紧密的膜结合信号）赋予GagPol质膜靶向，但Fyn（10）GagPol不产生病毒颗粒。粒子出芽的缺陷不能通过引入PTAP基序来挽救，PTAP基序负责病毒粒子出芽的后期阶段。相反，电子显微镜表明GagPR的出芽缺陷发生在颗粒形态发生的早期阶段。我们的数据，这是与以前的观察一致，证明了GagPol在膜结合和粒子组装的缺陷。
The GagPol protein of HIV-1 harbors viral enzymes, such as protease (PR), reverse transcriptase, and integrase, that are all crucial for virion infectivity. Previous studies have suggested that expression of GagPol alone does not produce viral particles and that the budding defect is caused by the presence of the Pol region. However, it has remained unknown why GagPol fails to produce viral particles. We show here that HIV-1 GagPol is incapable of membrane binding and subsequent particle assembly. Our confocal data indicated that, despite full N-myristoylation, GagPol protein failed to target plasma membrane with diffuse distribution in the cytoplasm. Membrane flotation analysis confirmed these findings. Progressive C-terminal truncation of GagPol to give GagPR allowed for plasma membrane targeting but still not for particle production. Conversely, the C-terminal addition of a noncognate protein, such as ß-galactosidase or 4 tandem GFP, to Gag impaired the membrane affinity, indicating that the Pol region, a large extension to Gag, inhibits membrane binding in the context of GagPol. The addition of the 10 N-terminal amino acids of Fyn kinase [Fyn(10)], a tight membrane-binding signal, conferred plasma membrane targeting on GagPol, but the Fyn(10)GagPol did not produce viral particles. The defect in particle budding was not rescued by the introduction of the PTAP motif, which is responsible for a late stage of viral particle budding. Rather, electron microscopy suggested that the budding defect of GagPR occurred at an early stage of particle morphogenesis. Our data, which were consistent with previous observations, demonstrate the defects of GagPol in membrane binding and particle assembly.
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DOI：
10.1083/jcb.200609050
发表时间：
2007-04-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Deneka M;Pelchen-Matthews A;Byland R;Ruiz-Mateos E;Marsh M
通讯作者：
Marsh M
DOI：
10.1073/pnas.86.15.5781
发表时间：
1989-08-01
影响因子：
11.1
作者：
GOTTLINGER, HG;SODROSKI, JG;HASELTINE, WA
通讯作者：
HASELTINE, WA
影响因子：
30.3
作者：
Jouve, Mabel;Sol-Foulon, Nathalie;Benaroch, Philippe
通讯作者：
Benaroch, Philippe
影响因子：
5.4
作者：
Dalton, Amanda K.;Ako-Adjei, Danso;Vogt, Volker M.
通讯作者：
Vogt, Volker M.
影响因子：
5.4
作者：
Finzi, Andres;Orthwein, Alexandre;Cohen, Eric A.
通讯作者：
Cohen, Eric A.