Assembly of rubella virus structural proteins into virus-like particles in transfected cells.
We have developed a stably transfected CHO cell line (CHO24S) that expresses the three structural proteins of rubella virus (RV). RV proteins C (capsid), E2, and E1 are secreted from CHO24S cells in the form of RV-like particles (RLPs) which form by budding into the cisterna of the Golgi complex. RLPs resemble RV virions in their size and morphology and have an identical buoyant density when purified on sucrose gradients. Release of RLPs into the medium was found to be dependent upon the E1 cytoplasmic tail since deletion or substitution of this domain with the same region from vesicular stomatitis virus G protein abrogated release of RV proteins from transfected cells. These results indicate that the RV 40S genomic RNA is not required for efficient particle assembly. Therefore, RLPs may serve as a convenient source of RV antigen for use in diagnostic assays and as an alternative to live attenuated vaccine strains.
Division of Cellular and Molecular Medicine, University of California at San Diego, La Jolla 92093-0651.
T C
TC
Hobman
M L
ML
Lundstrom
C A
CA
Mauracher
L
L
Woodward
S
S
Gillam
M G
MG
Farquhar
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D016466:CHO Cells; D002213:Capsid / Q000648:ultrastructure; D006224:Cricetinae; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009024:Morphogenesis; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins; D012412:Rubella virus / Q000276:immunology / Q000648:ultrastructure*; D013329:Structure-Activity Relationship; D014162:Transfection; D014614:Vaccines, Synthetic; D014764:Viral Proteins / Q000276:immunology; D014771:Virion / Q000648:ultrastructure
D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014764:Viral Proteins