Membrane-anchored incorporation of a foreign protein in recombinant influenza virions.
The RNA polymerase I system for in vivo synthesis of recombinant influenza vRNA molecules was used for the expression of a chimeric protein, consisting of the 341-amino-acid ectodomain of the glycoprotein E2 of classical swine fever virus and the 37-amino-acid C-terminal membrane anchor of the influenza virus hemagglutinin (HA). During infection with an influenza A helper virus the amplified pseudo-viral RNA was packaged into progeny virions together with influenza vRNA segments. The foreign fusion protein E2-HA was shown to be physically incorporated into the viral envelope. Incorporation of a third major glycoprotein into the envelope did not affect biological functions of HA and neuraminidase that are required for the generation of infectious virus particles. Based on mutational analyses of the cytoplasmic tail of E2-HA fusion proteins three modes of interaction during virus budding have been observed: nonspecific low-level incorporation (truncated tails), specific full-level incorporation (wild-type amino acid sequence or minor variations of it), and exclusion from incorporation (elongated tails).
Zhou
Y
Y
Institut für Mikrobiologie und Molekularbiologie, Universität Giessen, Germany.
König
M
M
Hobom
G
G
Neumeier
E
E
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D006692:Classical Swine Fever Virus; D003593:Cytoplasm / Q000821:virology; D004285:Dogs; D005822:Genetic Vectors / Q000235:genetics*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014771:Virion / Q000235:genetics; D019065:Virus Assembly / Q000235:genetics*
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus