CiTCoM, CNRS, UMR 8038, Université de Paris, Paris, France.
Biomol NMR Assign. 2021 Oct;15(2):267-271. doi: 10.1007/s12104-021-10016-9. Epub 2021 Mar 22.
During the maturation of the HIV-1 particle, the Gag polyprotein is cleaved by the viral protease into several proteins: matrix (MA), capsid (CA), spacer peptide 1 (SP1), nucleocapsid (NC), spacer peptide 2 (SP2) and p6. After cleavage, these proteins rearrange to form infectious viral particles. The final cleavage by the protease occurs between CA and SP1 and is the limiting step for the maturation of the particle. The CA-SP1 junction is the target of HIV-1 maturation inhibitors. CA is responsible for the formation of the viral capsid which protects the viral RNA inside. The SP1 domain is essential for viral assembly and infectivity, it is flexible and in helix-coil equilibrium. The presence of NC allows the SP1 domain to be less dynamic. The perturbation of the natural coil-helix equilibrium to helix interferes with protease cleavage and leads to non-completion of viral maturation. In this work, two mutations, W316A and M317A, that abolish the oligomerization of CA were introduced into the protein. The HIV-1 CA-SP1-NC which contains the C-terminal monomeric mutant of CA, SP1 and NC was produced to study the mechanism of action of HIV-1 maturation inhibitors. Here we report the backbone assignment of the protein CA-SP1-NC. These results will be useful to study the interaction between HIV-1 Gag and HIV-1 maturation inhibitors.
在 HIV-1 颗粒成熟过程中，Gag 多聚蛋白被病毒蛋白酶切割成几种蛋白质：基质（MA）、衣壳（CA）、间隔肽 1（SP1）、核衣壳（NC）、间隔肽 2（SP2）和 p6。切割后，这些蛋白质重新排列形成感染性病毒颗粒。最后由蛋白酶在 CA 和 SP1 之间切割，这是颗粒成熟的限速步骤。CA-SP1 连接处是 HIV-1 成熟抑制剂的靶标。CA 负责形成保护病毒 RNA 内部的病毒衣壳。SP1 结构域对于病毒组装和感染性至关重要，它是灵活的，处于螺旋-卷曲平衡状态。NC 的存在使 SP1 结构域的动态性降低。天然螺旋-卷曲平衡向螺旋的扰动会干扰蛋白酶的切割，导致病毒成熟不完全。在这项工作中，引入了两个突变 W316A 和 M317A，它们使 CA 的寡聚化作用丧失。产生含有 CA、SP1 和 NC 的 C 端单体突变体的 HIV-1 CA-SP1-NC 来研究 HIV-1 成熟抑制剂的作用机制。在这里，我们报告了蛋白 CA-SP1-NC 的骨架分配。这些结果将有助于研究 HIV-1 Gag 与 HIV-1 成熟抑制剂之间的相互作用。
