Evidence that the respiratory syncytial virus polymerase complex associates with lipid rafts in virus-infected cells: a proteomic analysis

• Published in: Virology (New York, N.Y.) N.Y.), 2004-12, Vol.330 (1), p.147-157
• Main Authors: McDonald, Terence P., Pitt, Andrew R., Brown, Gaie, Rixon, Helen W. McL, Sugrue, Richard J.
• Format: Article
• Language: English
• Subjects: 2D-nLC-MSMS; 60 APPLIED LIFE SCIENCES; Amino Acid Sequence; Cell Line, Tumor; DNA-Directed RNA Polymerases - chemistry; DNA-Directed RNA Polymerases - metabolism; ENZYME ACTIVITY; Humans; INTERACTIONS; Lipid raft; LIPIDS; Mass Spectrometry; MASS SPECTROSCOPY; Membrane Microdomains - virology; MEMBRANES; MICROSCOPY; Molecular Sequence Data; Peptide Fragments - chemistry; PROTEINS; Proteomics; Respiratory syncytial virus; Respiratory Syncytial Virus, Human - enzymology; Virus polymerase; VIRUSES
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2004.09.034

The interaction between the respiratory syncytial virus (RSV) polymerase complex and lipid rafts was examined in HEp2 cells. Lipid-raft membranes were prepared from virus-infected cells and their protein content was analysed by Western blotting and mass spectrometry. This analysis revealed the presence of the N, P, L, M2-1 and M proteins. However, these proteins appeared to differ from one another in their association with these structures, with the M2-1 protein showing a greater partitioning into raft membranes compared to that of the N, P or M proteins. Determination of the polymerase activity profile of the gradient fractions revealed that 95% of the detectable viral enzyme activity was associated with lipid-raft membranes. Furthermore, analysis of virus-infected cells by confocal microscopy suggested an association between these proteins and the raft-lipid, GM1. Together, these results provide evidence that the RSV polymerase complex is able to associate with lipid rafts in virus-infected cells.