Intrinsic cytoskeleton-dependent clustering of influenza virus M2 protein with hemagglutinin assessed by FLIM-FRET.
The hemagglutinin (HA) of influenza virus organizes the virus bud zone, a domain of the plasma membrane enriched in raft lipids. Using fluorescence lifetime imaging microscopy-fluorescence resonance energy transfer (FLIM-FRET), a technique that detects close colocalization of fluorescent proteins in transfected cells, we show that the viral proton channel M2 clusters with HA but not with a marker for inner leaflet rafts. The FRET signal between M2 and HA depends on the raft-targeting signals in HA and on an intact actin cytoskeleton. We conclude that M2 contains an intrinsic signal that targets the protein to the viral bud zone, which is organized by raft-associated HA and by cortical actin.
Thaa
Bastian
B
Department of Immunology and Molecular Biology, Veterinary Faculty, Free University, Berlin, Germany.
Herrmann
Andreas
A
Veit
Michael
M
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003599:Cytoskeleton / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006388:Hemagglutinins / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D014763:Viral Matrix Proteins / Q000378:metabolism*; D000086082:Viroporin Proteins
D006388:Hemagglutinins; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins