Cryo-ET of Env on intact HIV virions reveals structural variation and positioning on the Gag lattice

HIV-1 Env mediates viral entry into host cells and is the sole target for neutralizing antibodies. However, Env structure and organization in its native virion context has eluded detailed characterization. Here, we used cryo-electron tomography to analyze Env in mature and immature HIV-1 particles. Immature particles showed distinct Env positioning relative to the underlying Gag lattice, providing insights into long-standing questions about Env incorporation. A 9.1-A sub-tomogram-averaged reconstruction of virion-bound Env in conjunction with structural mass spectrometry revealed unexpected features, including a variable central core of the gp41 subunit, heterogeneous glycosylation between protomers, and a flexible stalk that allows Env tilting and variable exposure of neutralizing epitopes. Together, our results provide an integrative understanding of HIV assembly and structural variation in Env antigen presentation.

Automated summary

This study used cryo-electron tomography to analyze Env in HIV-1 particles and revealed distinct positioning of Env in immature particles relative to the underlying Gag lattice. Additionally, unexpected structural features of virion-bound Env, such as a variable central core and heterogeneous glycosylation, were discovered. These findings provide a comprehensive understanding of HIV assembly and structural variation in Env antigen presentation.