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-Structure paper
タイトル | HIV-1 Intasomes Assembled with Excess Integrase C-Terminal Domain Protein Facilitate Structural Studies by Cryo-EM and Reveal the Role of the Integrase C-Terminal Tail in HIV-1 Integration. |
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ジャーナル・号・ページ | Viruses, Vol. 16, Issue 7, Year 2024 |
掲載日 | 2024年7月20日 |
![]() | Min Li / Zhen Li / Xuemin Chen / Yanxiang Cui / Alan N Engelman / Robert Craigie / ![]() ![]() |
PubMed 要旨 | Retroviral integration is mediated by intasome nucleoprotein complexes wherein a pair of viral DNA ends are bridged together by a multimer of integrase (IN). Atomic-resolution structures of HIV-1 ...Retroviral integration is mediated by intasome nucleoprotein complexes wherein a pair of viral DNA ends are bridged together by a multimer of integrase (IN). Atomic-resolution structures of HIV-1 intasomes provide detailed insights into the mechanism of integration and inhibition by clinical IN inhibitors. However, previously described HIV-1 intasomes are highly heterogeneous and have the tendency to form stacks, which is a limiting factor in determining high-resolution cryo-EM maps. We have assembled HIV-1 intasomes in the presence of excess IN C-terminal domain protein, which was readily incorporated into the intasomes. The purified intasomes were largely homogeneous and exhibited minimal stacking tendencies. The cryo-EM map resolution was further improved to 2.01 Å, which will greatly facilitate structural studies of IN inhibitor action and drug resistance mechanisms. The C-terminal 18 residues of HIV-1 IN, which are critical for virus replication and integration in vitro, have not been well resolved in previous intasome structures, and its function remains unclear. We show that the C-terminal tail participates in intasome assembly, resides within the intasome core, and forms a small alpha helix (residues 271-276). Mutations that disrupt alpha helix integrity impede IN activity in vitro and disrupt HIV-1 infection at the step of viral DNA integration. |
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手法 | EM (単粒子) |
解像度 | 2.01 Å |
構造データ | EMDB-45364, PDB-9c9m: |
化合物 | ![]() ChemComp-MG: ![]() ChemComp-ZN: ![]() ChemComp-DLU: ![]() ChemComp-HOH: |
由来 |
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![]() | VIRAL PROTEIN / HIV-1 / integrase / nucleoprotein complexes / INSTI / intasome / CryoEM |