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Electron tomography visualization of HIV-1 fusion with target cells using fusion inhibitors to trap the pre-hairpin intermediate

Ladinsky, Mark S. and Gnanapragasam, Priyanthi N. P. and Yang, Zhi and West, Anthony P. and Kay, Michael S. and Bjorkman, Pamela J. (2020) Electron tomography visualization of HIV-1 fusion with target cells using fusion inhibitors to trap the pre-hairpin intermediate. eLife, 9 . Art. No. e58411. ISSN 2050-084X. PMCID PMC7394545. https://resolver.caltech.edu/CaltechAUTHORS:20200501-113540872

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Abstract

Fusion of HIV-1 with the membrane of its target cell, an obligate first step in virus infectivity, is mediated by binding of the viral envelope (Env) spike protein to its receptors, CD4 and CCR5/CXCR4, on the cell surface. The process of viral fusion appears to be fast compared with viral egress and has not been visualized by EM. To capture fusion events, the process must be curtailed by trapping Env-receptor binding at an intermediate stage. We have used fusion inhibitors to trap HIV-1 virions attached to target cells by Envs in an extended pre-hairpin intermediate state. Electron tomography revealed HIV-1 virions bound to TZM-bl cells by 2–4 narrow spokes, with slightly more spokes present when evaluated with mutant virions that lacked the Env cytoplasmic tail. These results represent the first direct visualization of the hypothesized pre-hairpin intermediate of HIV-1 Env and improve our understanding of Env-mediated HIV-1 fusion and infection of host cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.7554/eLife.58411DOIArticle
https://doi.org/10.1101/2020.04.29.068775DOIDiscussion Paper
https://caltech.box.com/s/b1rxyfr28ti75gl1q2qqs0lq87qvcz0qRelated ItemData/Code
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc7394545/PubMed CentralArticle
ORCID:
AuthorORCID
Ladinsky, Mark S.0000-0002-1036-3513
Yang, Zhi0000-0001-8680-3784
Kay, Michael S.0000-0003-3186-9684
Bjorkman, Pamela J.0000-0002-2277-3990
Alternate Title:Electron tomography visualization of HIV-1 fusion with target cells using inhibitors to trap the prehairpin intermediate
Additional Information:© 2020 Ladinsky et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: 29 April 2020; Accepted: 21 July 2020; Published: 22 July 2020. The authors thank Annie Lynch for assistance with initial experiments, Erica Lee, Jennifer Keeffe, and the Beckman Institute Protein Expression Center at Caltech for preparing T1249-Fc, D5 IgG, and Z004 IgG, Sarah Apple and Nicholas Francis for preparing CPT31, Magnus Hoffmann for suggestions, and members of the Bjorkman and Kay laboratories for helpful discussions and critical review of the manuscript. This work was supported by the National Institute of General Medical Sciences (2 P50 AI150464) (to MSK and PJB). We thank the Caltech Kavli Nanoscience Institute for maintenance of the TF-30 electron microscope. Competing interests: Pamela J Bjorkman: Reviewing editor, eLife. The other authors declare that no competing interests exist. Author contributions: Mark S Ladinsky, Conceptualization, Data curation, Formal analysis, Supervision, Funding acquisition, Visualization, Methodology, Writing - original draft, Writing - review and editing; Priyanthi NP Gna- napragasam, Conceptualization, Resources, Data curation, Formal analysis, Visualization, Methodol- ogy, Writing - original draft, Writing - review and editing; Zhi Yang, Resources, Data curation, Methodology, Writing - review and editing; Anthony P West, Data curation, Formal analysis, Meth- odology; Michael S Kay, Conceptualization, Resources, Formal analysis, Methodology, Writing - review and editing; Pamela J Bjorkman, Conceptualization, Resources, Formal analysis, Supervision, Funding acquisition, Writing - original draft, Writing - review and editing. Data availability: Raw datasets are freely available upon request. Interested parties should contact ladinsky@caltech.edu, and we will place requested datasets onto an externally accessible Caltech Box Server. Requestors will then be provided with a direct URL link from which they can download the files at their convenience.
Group:Kavli Nanoscience Institute
Funders:
Funding AgencyGrant Number
NIH2 P50 AI150464
National Institute of General Medical SciencesUNSPECIFIED
PubMed Central ID:PMC7394545
Record Number:CaltechAUTHORS:20200501-113540872
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200501-113540872
Official Citation:Electron tomography visualization of HIV-1 fusion with target cells using inhibitors to trap the prehairpin intermediate. eLife 2020; 9:e58411; DOI: 10.7554/eLife.58411
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:102968
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 May 2020 19:16
Last Modified:06 Aug 2020 21:41

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