Synchronized HIV assembly by tunable PIP2 changes reveals PIP2 requirement for stable Gag anchoring

Abstract
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Abstract

HIV-1 assembles at the plasma membrane (PM) of infected cells. PM association of the main structural protein Gag depends on its myristoylated MA domain and PM PI(4,5)P₂. Using a novel chemical biology tool that allows rapidly tunable manipulation of PI(4,5)P₂ levels in living cells, we show that depletion of PI(4,5)P₂ completely prevents Gag PM targeting and assembly site formation. Unexpectedly, PI(4,5)P₂ depletion also caused loss of pre-assembled Gag lattices from the PM. Subsequent restoration of PM PI(4,5)P₂ reinduced assembly site formation even in the absence of new protein synthesis, indicating that the dissociated Gag molecules remained assembly competent. These results reveal an important role of PI(4,5)P₂ for HIV-1 morphogenesis beyond Gag recruitment to the PM and suggest a dynamic equilibrium of Gag-lipid interactions. Furthermore, they establish an experimental system that permits synchronized induction of HIV-1 assembly leading to induced production of infectious virions by targeted modulation of Gag PM targeting.

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Frauke Mücksch

Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-0132-5101
Vibor Laketa

Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany

Competing interests
No competing interests declared.
Barbara Müller

Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-5726-5585
Carsten Schultz

Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany

Competing interests
Carsten Schultz, C. Schultz is a shareholder of the company SiChem, which distributes rCDS.
Hans-Georg Kräusslich

Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany

For correspondence
hans-georg.kraeusslich@med.uni-heidelberg.de

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-8756-329X

Funding

Deutsche Forschungsgemeinschaft (TRR 83 project 14)

Hans-Georg Kräusslich

Deutsche Forschungsgemeinschaft (TRR 83 project 2)

Carsten Schultz

Deutsche Forschungsgemeinschaft (SFB 1129 project 5)

Hans-Georg Kräusslich

Deutsche Forschungsgemeinschaft (SFB 1129 project 6)

Barbara Müller

Deutsche Forschungsgemeinschaft (Excellence Cluster CellNetworks Exc81)

Barbara Müller
Hans-Georg Kräusslich

Deutsches Zentrum für Infektionsforschung (Project 7.5 TTU HIV)

Hans-Georg Kräusslich

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.