Murine leukemia virus nucleocapsid mutant particles lacking viral RNA encapsidate ribosomes.
A single retroviral protein, termed Gag, is sufficient for assembly of retrovirus-like particles in mammalian cells. Gag normally selects the genomic RNA of the virus with high specificity; the nucleocapsid (NC) domain of Gag plays a crucial role in this selection process. However, encapsidation of the viral RNA is completely unnecessary for particle assembly. We previously showed that mutant murine leukemia virus (MuLV) particles that lack viral RNA because of a deletion in the cis-acting packaging signal ("Psi") in the genomic RNA compensate for the loss of the viral RNA by incorporating cellular mRNA. The RNA in wild-type and Psi- particles was also found to be necessary for virion core structure. In the present work, we explored the role of RNA in MuLV particles that lack genomic RNA because of mutations in the NC domain of Gag. Using a fluorescent dye assay, we observed that NC mutant particles contain the same amount of RNA that wild-type virions do. Surprisingly enough, these particles contained large amounts of rRNAs. Furthermore, ribosomal proteins were detected by immunoblotting, and ribosomes were observed inside the particles by electron microscopy. The biological significance of the presence of ribosomes in NC mutant particles lacking genomic RNA is discussed.
Muriaux
Delphine
D
HIV Drug Resistance Program, and Image Analysis Laboratory. SAIC-Frederick, National Cancer Institute-Frederick, Frederick, Maryland 21702-1201, USA. dmuriaux@nifcrf.gov
Mirro
Jane
J
Nagashima
Kunio
K
Harvin
Demetria
D
Rein
Alan
A
D000818:Animals; D002460:Cell Line; D016679:Genome, Viral; D006801:Humans; D015151:Immunoblotting; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008854:Microscopy, Electron; D009154:Mutation; D019251:Nucleocapsid / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*; D019065:Virus Assembly
D012335:RNA, Ribosomal; D012367:RNA, Viral