Abstract
Retrovirus budding requires short peptide motifs (late domains) located within the viral Gag protein that function by recruiting cellular factors. The YPXnL late domains of HIV and other lentiviruses recruit the protein ALIX (also known as AIP1), which also functions in vesicle formation at the multivesicular body and in the abscission stage of cytokinesis. Here, we report the crystal structures of ALIX in complex with the YPXnL late domains from HIV-1 and EIAV. The two distinct late domains bind at the same site on the ALIX V domain but adopt different conformations that allow them to make equivalent contacts. Binding studies and functional assays verified the importance of key interface residues and revealed that binding affinities are tuned by context-dependent effects. These results reveal how YPXnL late domains recruit ALIX to facilitate virus budding and how ALIX can bind YPXnL sequences with both n = 1 and n = 3.
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Acknowledgements
We thank P. Bieniasz (Aaron Diamond AIDS Research Center) for kindly providing various HIV-1 proviral plasmids and H. Göttlinger (University of Massachusettes) for helpful discussions. This work was funded by US National Institutes of Health (NIH) grants GM082534 (C.P.H., W.I.S.) and AI051174 (W.I.S.). Portions of this research were performed at the Stanford Synchrotron Radiation Laboratory (SSRL), a national user facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research, and by the NIH, National Center for Research Resources, Biomedical Technology Program and National Institute of General Medical Sciences. The Center for Biomolecular Interactions Analysis at the University of Utah is funded in part by NIH grant 1S10RR016787-01 (D.G.M.).
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Zhai, Q., Fisher, R., Chung, HY. et al. Structural and functional studies of ALIX interactions with YPXnL late domains of HIV-1 and EIAV. Nat Struct Mol Biol 15, 43–49 (2008). https://doi.org/10.1038/nsmb1319
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DOI: https://doi.org/10.1038/nsmb1319
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