Architecture of a nascent viral fusion pore.
Enveloped viruses use specialized protein machinery to fuse the viral membrane with that of the host cell during cell invasion. In influenza virus, hundreds of copies of the haemagglutinin (HA) fusion glycoprotein project from the virus surface. Despite intensive study of HA and its fusion activity, the protein's modus operandi in manipulating viral and target membranes to catalyse their fusion is poorly understood. Here, the three-dimensional architecture of influenza virus-liposome complexes at pH 5.5 was investigated by electron cryo-tomography. Tomographic reconstructions show that early stages of membrane remodeling take place in a target membrane-centric manner, progressing from punctate dimples, to the formation of a pinched liposomal funnel that may impinge on the apparently unperturbed viral envelope. The results suggest that the M1 matrix layer serves as an endoskeleton for the virus and a foundation for HA during membrane fusion. Fluorescence spectroscopy monitoring fusion between liposomes and virions shows that leakage of liposome contents takes place more rapidly than lipid mixing at pH 5.5. The relation of 'leaky' fusion to the observed prefusion structures is discussed.
Lee
Kelly K
KK
Department of Medicinal Chemistry, University of Washington, Seattle, WA 98195-7610, USA. kklee@u.washington.edu
D020285:Cryoelectron Microscopy; D055032:Electron Microscope Tomography; D006388:Hemagglutinins / Q000032:analysis / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D008081:Liposomes / Q000378:metabolism* / Q000648:ultrastructure*; D008561:Membrane Fusion; D010713:Phosphatidylcholines / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014759:Viral Envelope Proteins / Q000032:analysis; D014760:Viral Fusion Proteins / Q000032:analysis / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
D006388:Hemagglutinins; D008081:Liposomes; D010713:Phosphatidylcholines; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; C017251:1,2-oleoylphosphatidylcholine