Encapsidation of heterologous RNAs by bacteriophage MS2 coat protein.
The RNA bacteriophages of E. coli specifically encapsidate a single copy of the viral genome in a protein shell composed mainly of 180 molecules of coat protein. Coat protein is also a translational repressor and shuts off viral replicase synthesis by interaction with a RNA stem-loop containing the replicase initiation codon. We wondered whether the translational operator also serves as the viral pac site, the signal which mediates the exclusive encapsidation of viral RNA by its interaction with coat protein. To test this idea we measured the ability of lacZ RNA fused to the translational operator to be incorporated into virus-like particles formed from coat protein expressed from a plasmid. The results indicate that the operator-lacZ RNA is indeed encapsidated and that nucleotide substitutions in the translational operator which reduce the tightness of the coat protein-operator interaction also reduce or abolish encapsidation of the hybrid RNA. When coat protein is expressed in excess compared to the operator-lacZ RNA, host RNAs are packaged as well. However, elevation of the level of operator-lacZ RNA relative to coat protein results in its selective encapsidation at the expense of cellular RNAs. Our results are consistent with the proposition that this single protein-RNA interaction accounts both for translational repression and viral genome encapsidation.
Pickett
G G
GG
Department of Cell Biology, University of New Mexico, School of Medicine and Cancer Center, Albuquerque 87131.
Peabody
D S
DS
D001483:Base Sequence; D002213:Capsid / Q000378:metabolism*; D004274:DNA, Recombinant; D017909:Levivirus / Q000378:metabolism*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009875:Operator Regions, Genetic; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism*; D012261:Ribonucleoproteins; D014779:Virus Replication
D004274:DNA, Recombinant; D046911:Macromolecular Substances; D012333:RNA, Messenger; D012261:Ribonucleoproteins