Abstract
Bacteriophage phi 6 has a segmented genome consisting of three pieces of double-stranded RNA (dsRNA). The viral procapsid is the structure that packages plus strands, synthesizes the complementary negative strands to form dsRNA, and then transcribes dsRNA to form plus-strand message. The minus-strand synthesis of a particular genomic segment is dependent on prior packaging of the other segments. The 5' end of the plus strand is necessary and sufficient for packaging, while the normal 3' end is necessary for synthesis of the negative strand. We have now investigated the ability of truncated RNA segments which lack the normal 3' end of the molecules to stimulate the synthesis of minus strands of the other segments. Fragments missing the normal 3' ends were able to stimulate the minus-strand synthesis of intact heterologous segments. Minus-strand synthesis of one intact segment could be stimulated by the presence of two truncated nonreplicating segments. The 5' fragments of each single-stranded genomic segment can compete with homologous full-length single-stranded genomic segments in minus-strand synthesis reactions, suggesting that there is a specific binding site in the procapsid for each segment.
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- Coplin D. L., Van Etten J. L., Koski R. K., Vidaver A. K. Intermediates in the biosynthesis of double-stranded ribonucleic acids of bacteriophage phi 6. Proc Natl Acad Sci U S A. 1975 Mar;72(3):849–853. doi: 10.1073/pnas.72.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Day L. A., Mindich L. The molecular weight of bacteriophage phi 6 and its nucleocapsid. Virology. 1980 Jun;103(2):376–385. doi: 10.1016/0042-6822(80)90196-8. [DOI] [PubMed] [Google Scholar]
- Emori Y., Iba H., Okada Y. Semi-conservative transcription of double-stranded RNA catalyzed by bacteriophage phi 6 RNA polymerase. J Biochem. 1980 Dec;88(6):1569–1575. doi: 10.1093/oxfordjournals.jbchem.a133131. [DOI] [PubMed] [Google Scholar]
- Ewen M. E., Revel H. R. In vitro replication and transcription of the segmented double-stranded RNA bacteriophage phi 6. Virology. 1988 Aug;165(2):489–498. doi: 10.1016/0042-6822(88)90593-4. [DOI] [PubMed] [Google Scholar]
- Ewen M. E., Revel H. R. RNA-protein complexes responsible for replication and transcription of the double-stranded RNA bacteriophage phi 6. Virology. 1990 Oct;178(2):509–519. doi: 10.1016/0042-6822(90)90348-u. [DOI] [PubMed] [Google Scholar]
- Gottlieb P., Strassman J., Bamford D. H., Mindich L. Production of a polyhedral particle in Escherichia coli from a cDNA copy of the large genomic segment of bacteriophage phi 6. J Virol. 1988 Jan;62(1):181–187. doi: 10.1128/jvi.62.1.181-187.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottlieb P., Strassman J., Frucht A., Qiao X. Y., Mindich L. In vitro packaging of the bacteriophage phi 6 ssRNA genomic precursors. Virology. 1991 Apr;181(2):589–594. doi: 10.1016/0042-6822(91)90892-f. [DOI] [PubMed] [Google Scholar]
- Gottlieb P., Strassman J., Qiao X. Y., Frucht A., Mindich L. In vitro replication, packaging, and transcription of the segmented double-stranded RNA genome of bacteriophage phi 6: studies with procapsids assembled from plasmid-encoded proteins. J Bacteriol. 1990 Oct;172(10):5774–5782. doi: 10.1128/jb.172.10.5774-5782.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottlieb P., Strassman J., Qiao X., Frilander M., Frucht A., Mindich L. In vitro packaging and replication of individual genomic segments of bacteriophage phi 6 RNA. J Virol. 1992 May;66(5):2611–2616. doi: 10.1128/jvi.66.5.2611-2616.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mindich L. Bacteriophage phi 6: a unique virus having a lipid-containing membrane and a genome composed of three dsRNA segments. Adv Virus Res. 1988;35:137–176. doi: 10.1016/s0065-3527(08)60710-1. [DOI] [PubMed] [Google Scholar]
- Mindich L., Qiao X., Onodera S., Gottlieb P., Strassman J. Heterologous recombination in the double-stranded RNA bacteriophage phi 6. J Virol. 1992 May;66(5):2605–2610. doi: 10.1128/jvi.66.5.2605-2610.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olkkonen V. M., Gottlieb P., Strassman J., Qiao X. Y., Bamford D. H., Mindich L. In vitro assembly of infectious nucleocapsids of bacteriophage phi 6: formation of a recombinant double-stranded RNA virus. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9173–9177. doi: 10.1073/pnas.87.23.9173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olkkonen V. M., Ojala P. M., Bamford D. H. Generation of infectious nucleocapsids by in vitro assembly of the shell protein on to the polymerase complex of the dsRNA bacteriophage phi 6. J Mol Biol. 1991 Apr 5;218(3):569–581. doi: 10.1016/0022-2836(91)90702-8. [DOI] [PubMed] [Google Scholar]
- Patton J. T. Evidence for equimolar synthesis of double-strand RNA and minus-strand RNA in rotavirus-infected cells. Virus Res. 1990 Nov;17(3):199–208. doi: 10.1016/0168-1702(90)90065-j. [DOI] [PubMed] [Google Scholar]
- Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
- Vidaver A. K., Koski R. K., Van Etten J. L. Bacteriophage phi6: a Lipid-Containing Virus of Pseudomonas phaseolicola. J Virol. 1973 May;11(5):799–805. doi: 10.1128/jvi.11.5.799-805.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- Zou S., Brown E. G. Identification of sequence elements containing signals for replication and encapsidation of the reovirus M1 genome segment. Virology. 1992 Feb;186(2):377–388. doi: 10.1016/0042-6822(92)90003-8. [DOI] [PubMed] [Google Scholar]