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Biochemical characterization of rous sarcoma virus MA protein interaction with membranes

Amanda K Dalton1, Paul S Murray, Diana Murray

  • 1Department of Molecular Biology and Genetics, Cornell University, 360 Biotechnology Building, Ithaca, NY 14853, USA.

Journal of Virology
|April 29, 2005
PubMed

Insights

Rous sarcoma virus MA protein binds to negatively charged lipids electrostatically. MA domain clustering, not intrinsic affinity, drives virus assembly at the host cell membrane.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • The MA domain of retroviral Gag proteins is crucial for virus assembly at the host cell membrane.
  • The precise biochemical mechanisms governing MA-membrane interactions remain poorly understood.

Purpose of the Study:

  • To investigate the in vitro biophysical properties of Rous sarcoma virus (RSV) MA domain-membrane interactions.
  • To elucidate the role of electrostatic forces and lipid composition in MA-membrane binding.

Main Methods:

  • In vitro liposome flotation assays using purified MA proteins.
  • Liposomes with defined lipid compositions were employed.
  • Computational modeling was used to support experimental findings.

Main Results:

  • RSV MA protein association with liposomes is electrostatic, requiring negatively charged lipids.
  • A mutant MA protein defective in vivo also failed to bind liposomes.
  • Monomeric MA showed weak membrane affinity, while artificially dimerized MA exhibited significantly higher binding affinity.

Conclusions:

  • The intrinsic affinity of monomeric RSV MA for negatively charged membranes is insufficient for efficient plasma membrane association during assembly.
  • Clustering of MA domains, facilitated by Gag-Gag interactions during virus assembly, likely drives the strong membrane association observed in vivo.

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