Definition of the interacting interfaces of Apobec3G and HIV-1 Vif using MAPPIT mutagenesis analysis
Definition of the interacting interfaces of Apobec3G and HIV-1 Vif using MAPPIT mutagenesis analysis
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DOI：
10.1093/nar/gkp1154
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发表时间：
2010-04-01
影响因子：
14.9
通讯作者：
Tavernier, Jan
中科院分区：
文献类型：
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作者：
Lavens, Delphine;Peelman, Frank;Tavernier, Jan
宿主限制因子Apobec 3G是一种胞苷脱氨酶，可掺入HIV-1病毒体并干扰病毒复制。HIV-1辅助蛋白Vif通过靶向Apobec 3G进行蛋白酶体降解来破坏Apobec 3G。我们提出了一个模型，其中Apobec 3G N-末端结构域对称相互作用，通过一个头对头的接口，包含残基122 RLYYFW 127。为了验证该模型并表征Apobec 3G-Apobec 3G和Apobec 3G-Vif相互作用，使用哺乳动物蛋白质-蛋白质相互作用陷阱双杂交技术。头对头界面中的突变消除了Apobec 3G-Apobec 3G相互作用。抑制Apobec 3G-Apobec 3G结合的所有突变也抑制Apobec 3G-Vif相互作用，表明头对头界面在与Vif的相互作用中起重要作用。只有D128 K、P129 A和T32 Q突变特异性地影响Apobec 3G-Vif关联。在我们的模型中，D128，P129和T32聚集在头-头界面的边缘，可能形成由两个Apobec 3G分子组成的Vif结合位点。我们建议Vif要么在Apobec 3G头对头界面结合，要么与RNA稳定的Apobec 3G寡聚体结合。
The host restriction factor Apobec3G is a cytidine deaminase that incorporates into HIV-1 virions and interferes with viral replication. The HIV-1 accessory protein Vif subverts Apobec3G by targeting it for proteasomal degradation. We propose a model in which Apobec3G N-terminal domains symmetrically interact via a head-to-head interface containing residues 122 RLYYFW 127. To validate this model and to characterize the Apobec3G-Apobec3G and the Apobec3G-Vif interactions, the mammalian protein-protein interaction trap two-hybrid technique was used. Mutations in the head-to-head interface abrogate the Apobec3G-Apobec3G interaction. All mutations that inhibit Apobec3G-Apobec3G binding also inhibit the Apobec3G-Vif interaction, indicating that the head-to head interface plays an important role in the interaction with Vif. Only the D128K, P129A and T32Q mutations specifically affect the Apobec3G-Vif association. In our model, D128, P129 and T32 cluster at the edge of the head-to-head interface, possibly forming a Vif binding site composed of two Apobec3G molecules. We propose that Vif either binds at the Apobec3G head-to-head interface or associates with an RNA-stabilized Apobec3G oligomer.