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Citron kinase enhances ubiquitination of HIV-1 Gag protein and intracellular HIV-1 budding

Archives of Virology volume 161, pages 2441–2448 (2016)Cite this article

Abstract

Assembly and budding of human immunodeficiency virus type 1 (HIV-1) particles is a complex process involving a number of host proteins. We have previously reported that the RhoA effector citron kinase enhances HIV-1 production. However, the underlying mechanism is not clear. In this study, we found that citron kinase interacted with HIV-1 Gag protein via its zinc finger and leucine zipper domains. Electron microscopy analysis revealed that citron kinase induced viral particle assembly in multivesicular bodies (MVBs). Citron kinase enhanced ubiquitination of HIV-1 Gag protein. Knockdown of Nedd4L, a member of the HECT ubiquitin E3 ligase family, partly decreased the ability of citron kinase to enhance HIV-1 production and reduced ubiquitination of HIV-1 Gag. Interestingly, the function of citron kinase to promote HIV-1 budding was severely impaired when endogenous ALIX was knocked down. Overexpression of the AAA-type ATPase VPS4 eliminated citron-kinase-mediated enhancement of HIV-1 production. Our results suggest that citron kinase interacts with HIV-1 Gag and enhances HIV-1 production by promoting Gag ubiquitination and inducing viral release via the MVB pathway.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (31200136), 973 Program (2012CB911102 CS) and National Mega-Project for Infectious Disease (2013ZX1000 4601-002 CS).

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    Affiliations

    1. Institute of Medicinal Biotechnology, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China

      Jiwei Ding, Jianyuan Zhao, Zeyun Mi & Shan Cen

    2. Center for Biological Imaging, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

      Lei Sun

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    1. Jiwei Ding
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    2. Jianyuan Zhao
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    Correspondence to Shan Cen.

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    Funding

    This study was funded by the National Natural Science Foundation of China (grant number: 31200136), 973 Program (grant number: 2012CB911102 CS) and National Mega-Project for Infectious Disease (grant number: 2013ZX1000 4601-002 CS).

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    All authors declare that they have no conflict of interest.

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    This article does not contain any study with human participants or animals performed by any of the authors.

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    J. Ding and J. Zhao contributes equally to the work.

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    Cite this article

    Ding, J., Zhao, J., Sun, L. et al. Citron kinase enhances ubiquitination of HIV-1 Gag protein and intracellular HIV-1 budding. Arch Virol 161, 2441–2448 (2016). https://doi.org/10.1007/s00705-016-2933-5

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    • DOI: https://doi.org/10.1007/s00705-016-2933-5

    Keywords

    • HEK293T Cell
    • Leucine Zipper Domain
    • Citron Kinase
    • p24CA ELISA
    • Late Endosomal Pathway