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Live-cell observation of cytosolic HIV-1 assembly onset reveals RNA-interacting Gag oligomers.
Hendrix, Jelle; Baumgärtel, Viola; Schrimpf, Waldemar; Ivanchenko, Sergey; Digman, Michelle A; Gratton, Enrico; Kräusslich, Hans-Georg; Müller, Barbara; Lamb, Don C.
Affiliation
  • Hendrix J; Physical Chemistry, Department of Chemistry, Ludwig Maximilian University of Munich, D-81377 Munich, Germany NanoSystems Initiative Munich (NIM), Ludwig Maximilian University of Munich, D-81377 Munich, Germany Munich Center for Integrated Protein Science (CiPSM), Ludwig Maximilian University of Muni
  • Baumgärtel V; Physical Chemistry, Department of Chemistry, Ludwig Maximilian University of Munich, D-81377 Munich, Germany NanoSystems Initiative Munich (NIM), Ludwig Maximilian University of Munich, D-81377 Munich, Germany Munich Center for Integrated Protein Science (CiPSM), Ludwig Maximilian University of Muni
  • Schrimpf W; Physical Chemistry, Department of Chemistry, Ludwig Maximilian University of Munich, D-81377 Munich, Germany NanoSystems Initiative Munich (NIM), Ludwig Maximilian University of Munich, D-81377 Munich, Germany Munich Center for Integrated Protein Science (CiPSM), Ludwig Maximilian University of Muni
  • Ivanchenko S; Physical Chemistry, Department of Chemistry, Ludwig Maximilian University of Munich, D-81377 Munich, Germany NanoSystems Initiative Munich (NIM), Ludwig Maximilian University of Munich, D-81377 Munich, Germany Munich Center for Integrated Protein Science (CiPSM), Ludwig Maximilian University of Muni
  • Digman MA; Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697 Development Biology Center Optical Biology Core Facility, University of California, Irvine, Irvine, CA 92697.
  • Gratton E; Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697 Development Biology Center Optical Biology Core Facility, University of California, Irvine, Irvine, CA 92697.
  • Kräusslich HG; Department of Infectious Diseases, Virology, University Hospital Heidelberg, D-69120 Heidelberg, Germany.
  • Müller B; Department of Infectious Diseases, Virology, University Hospital Heidelberg, D-69120 Heidelberg, Germany.
  • Lamb DC; Physical Chemistry, Department of Chemistry, Ludwig Maximilian University of Munich, D-81377 Munich, Germany NanoSystems Initiative Munich (NIM), Ludwig Maximilian University of Munich, D-81377 Munich, Germany Munich Center for Integrated Protein Science (CiPSM), Ludwig Maximilian University of Muni
J Cell Biol ; 210(4): 629-46, 2015 Aug 17.
Article in En | MEDLINE | ID: mdl-26283800
Assembly of the Gag polyprotein into new viral particles in infected cells is a crucial step in the retroviral replication cycle. Currently, little is known about the onset of assembly in the cytosol. In this paper, we analyzed the cytosolic HIV-1 Gag fraction in real time in live cells using advanced fluctuation imaging methods and thereby provide detailed insights into the complex relationship between cytosolic Gag mobility, stoichiometry, and interactions. We show that Gag diffuses as a monomer on the subsecond timescale with severely reduced mobility. Reduction of mobility is associated with basic residues in its nucleocapsid (NC) domain, whereas capsid (CA) and matrix (MA) domains do not contribute significantly. Strikingly, another diffusive Gag species was observed on the seconds timescale that oligomerized in a concentration-dependent manner. Both NC- and CA-mediated interactions strongly assist this process. Our results reveal potential nucleation steps of cytosolic Gag fractions before membrane-assisted Gag assembly.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: HIV-1 / Virus Assembly / Gag Gene Products, Human Immunodeficiency Virus Limits: Humans Language: En Journal: J Cell Biol Year: 2015 Type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: HIV-1 / Virus Assembly / Gag Gene Products, Human Immunodeficiency Virus Limits: Humans Language: En Journal: J Cell Biol Year: 2015 Type: Article