The cytoplasmic domain of alphavirus E2 glycoprotein contains a short linear recognition signal required for viral budding.
Intracellular alphavirus nucleocapsids express a binding site for the cytoplasmic domain of the viral E2 spike glycoprotein. This binding site is recognized by the anti-idiotype monoclonal antibody, F13. The monoclonal anti-anti-idiotype antibody, raised against F13 and designated 3G10, recognizes the carboxy-terminal eight residues of the E2 cytoplasmic domain in Semliki Forest virus (SFV), identifying this as the signal for nucleocapsid interaction. F13 binding to cells infected with SFV or a second alphavirus, Sindbis virus, is inhibited by a synthetic peptide corresponding to the entire 31 residue cytoplasmic domain (E2c), and also by a synthetic peptide corresponding to the eight residue epitope recognized by 3G10. Both E2c and the eight residue peptide inhibited viral budding in microinjection experiments and when conjugated to colloidal gold are bound specifically to nucleocapsids in infected cells. These results identify a short linear signal in the E2 cytoplasmic domain required for the interaction with nucleocapsids which leads to budding of at least two alphaviruses from infected cells.
Kail
M
M
Cell Biology Programme, European Molecular Biology Laboratory, Heidelberg, FRG.
Hollinshead
M
M
Ansorge
W
W
Pepperkok
R
R
Frank
R
R
Griffiths
G
G
Vaux
D
D
D000595:Amino Acid Sequence; D001665:Binding Sites; D002213:Capsid / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D008845:Microinjections; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000378:metabolism*; D012672:Semliki forest virus / Q000378:metabolism* / Q000502:physiology; D016415:Sequence Alignment; D012845:Sindbis Virus / Q000378:metabolism* / Q000502:physiology; D014617:Vacuoles / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism* / Q000648:ultrastructure; D014779:Virus Replication
D000939:Epitopes; D021382:Protein Sorting Signals; D014759:Viral Envelope Proteins; C064950:glycoprotein E2, Sindbis virus