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. 1980 Oct;32(1):295–312. doi: 10.1016/S0006-3495(80)84958-7

RNA-protein interactions in the assembly of tobacco mosaic virus.

P J Butler, G P Lomonossoff
PMCID: PMC1327308  PMID: 7248449

Abstract

Assembly of tobacco mosaic virus is initiated by the binding of a specific loop of the RNA into the central hole of the disk aggregate of protein subunits. Since the nucleation loop is located about five-sixths along the RNA molecule, subsequent elongation must be bidirectional. We have now measured the rates of elongation in the two directions by determining the lengths of RNA protected from nuclease digestion at different times and using either intact TMV rNA, or RNA with most of the longer tail removed. Comparison of the rates with the protein supplied as either a mixture of disks with A-protein (a mixture of less aggregated states) or just A-protein, shows that different mechanisms and protein aggregates are used for the most rapid growth. When disks are present, they add more rapidly along the longer RNA tail but do not appear to add directly on the shorter tail. In contrast, smaller aggregates (A-protein) can add at both ends of the rod, but do so more slowly. Mechanisms for these processes are discussed. Preliminary results on the binding of the specific hexanucleotide AAGAAG to the disk are given and compared with the known changes on binding nonspecific hexanucleotides or the trinucleotide AAG.

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Selected References

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  1. Butler P. J. Assembly of tobacco mosaic virus. Philos Trans R Soc Lond B Biol Sci. 1976 Nov 30;276(943):151–163. doi: 10.1098/rstb.1976.0106. [DOI] [PubMed] [Google Scholar]
  2. Butler P. J., Durham A. C., Klug A. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. IV. Control of mode of aggregation of tobacco mosaic virus protein by proton binding. J Mol Biol. 1972 Dec 14;72(1):1–18. doi: 10.1016/0022-2836(72)90063-0. [DOI] [PubMed] [Google Scholar]
  3. Butler P. J., Finch J. T. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. VII. Lengths of the growing rods during assembly into nucleoprotein with the viral RNA. J Mol Biol. 1973 Aug 25;78(4):637–649. doi: 10.1016/0022-2836(73)90285-4. [DOI] [PubMed] [Google Scholar]
  4. Butler P. J., Finch J. T., Zimmern D. Configuration of tobacco mosaic virus, RNA during virus assembly. Nature. 1977 Jan 20;265(5591):217–219. doi: 10.1038/265217a0. [DOI] [PubMed] [Google Scholar]
  5. Butler P. J., Klug A. Assembly of the particle of tobacco mosaic virus from RNA and disks of protein. Nat New Biol. 1971 Jan 13;229(2):47–50. doi: 10.1038/newbio229047a0. [DOI] [PubMed] [Google Scholar]
  6. Butler P. J., Klug A. Assembly of tobacco mosaic virus in vitro: effect of state of polymerization of the protein component. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2950–2953. doi: 10.1073/pnas.69.10.2950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Butler P. J., Klug A. The assembly of a virus. Sci Am. 1978 Nov;239(5):62–69. doi: 10.1038/scientificamerican1178-62. [DOI] [PubMed] [Google Scholar]
  8. Butler P. J., Lomonossoff G. P. Quantized incorporation of RNA during assembly of tobacco mosaic virus from protein disks. J Mol Biol. 1978 Dec 25;126(4):877–882. doi: 10.1016/0022-2836(78)90027-x. [DOI] [PubMed] [Google Scholar]
  9. Butler P. J. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. 8. Elongation of nucleoprotein rods of the virus RNA and protein. J Mol Biol. 1974 Jan 25;82(3):333–341. doi: 10.1016/0022-2836(74)90594-4. [DOI] [PubMed] [Google Scholar]
  10. Butler P. J. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. 8. Initial stages of assembly of nucleoprotein rods from virus RNA and the protein disks. J Mol Biol. 1974 Jan 25;82(3):343–353. doi: 10.1016/0022-2836(74)90595-6. [DOI] [PubMed] [Google Scholar]
  11. Butler P. J. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. VI. Assembly of the nucleoprotein rods of tobacco mosaic virus from the protein disks and RNA. J Mol Biol. 1972 Dec 14;72(1):25–35. doi: 10.1016/0022-2836(72)90065-4. [DOI] [PubMed] [Google Scholar]
  12. CASPAR D. L. ASSEMBLY AND STABILITY OF THE TOBACCO MOSAIC VIRUS PARTICLE. Adv Protein Chem. 1963;18:37–121. doi: 10.1016/s0065-3233(08)60268-5. [DOI] [PubMed] [Google Scholar]
  13. Champness J. N., Bloomer A. C., Bricogne G., Butler P. G., Klug A. The structure of the protein disk of tobacco mosaic virus to 5A resolution. Nature. 1976 Jan 1;259(5538):20–24. doi: 10.1038/259020a0. [DOI] [PubMed] [Google Scholar]
  14. Durham A. C., Klug A. Polymerization of tobacco mosaic virus protein and its control. Nat New Biol. 1971 Jan 13;229(2):42–46. doi: 10.1038/newbio229042a0. [DOI] [PubMed] [Google Scholar]
  15. Durham A. C. Structures and roles of the polymorphic forms of tobacco mosaic virus protein. I. Sedimentation studies. J Mol Biol. 1972 Jun 20;67(2):289–305. doi: 10.1016/0022-2836(72)90242-2. [DOI] [PubMed] [Google Scholar]
  16. FRAENKEL-CONRAT H., SINGER B. RECONSTITUTION OF TOBACCO MOSAIC VIRUS. IV. INHIBITION BY ENZYMES AND OTHER PROTEINS, AND USE OF POLYNUCLEOTIDES. Virology. 1964 Jul;23:354–362. doi: 10.1016/0042-6822(64)90257-0. [DOI] [PubMed] [Google Scholar]
  17. FRAENKEL-CONRAT H., SINGER B. Reconstitution of tobacco mosaic virus. III. Improved methods and the use of mixed nucleic acids. Biochim Biophys Acta. 1959 Jun;33(2):359–370. doi: 10.1016/0006-3002(59)90126-x. [DOI] [PubMed] [Google Scholar]
  18. Fraenkel-Conrat H., Williams R. C. RECONSTITUTION OF ACTIVE TOBACCO MOSAIC VIRUS FROM ITS INACTIVE PROTEIN AND NUCLEIC ACID COMPONENTS. Proc Natl Acad Sci U S A. 1955 Oct 15;41(10):690–698. doi: 10.1073/pnas.41.10.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fukuda M., Ohno T., Okada Y., Otsuki Y., Takebe I. Kinetics of biphasic reconstitution of tobacco mosaic virus in vitro. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1727–1730. doi: 10.1073/pnas.75.4.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Graham J., Butler P. J. Binding of oligonucleotides to the disk of tobacco-mosaic-virus protein. Eur J Biochem. 1979 Jan 15;93(2):333–337. doi: 10.1111/j.1432-1033.1979.tb12827.x. [DOI] [PubMed] [Google Scholar]
  21. Guilley H., Jonard G., Kukla B., Richards K. E. Sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA. Nucleic Acids Res. 1979 Apr;6(4):1287–1308. doi: 10.1093/nar/6.4.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Guilley H., Jonard G., Kukla B., Richards K. E. Sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA. Nucleic Acids Res. 1979 Apr;6(4):1287–1308. doi: 10.1093/nar/6.4.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jonard G., Richards K. E., Guilley H., Hirth L. Sequence from the assembly nucleation region of TMV RNA. Cell. 1977 Jul;11(3):483–493. doi: 10.1016/0092-8674(77)90066-6. [DOI] [PubMed] [Google Scholar]
  24. Jonathan P., Butler G., Durham A. C. Tobacco mosaic virus protein aggregation and the virus assembly. Adv Protein Chem. 1977;31:187–251. doi: 10.1016/s0065-3233(08)60219-3. [DOI] [PubMed] [Google Scholar]
  25. Lebeurier G., Nicolaieff A., Richards K. E. Inside-out model for self-assembly of tobacco mosaic virus. Proc Natl Acad Sci U S A. 1977 Jan;74(1):149–153. doi: 10.1073/pnas.74.1.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lomonossoff G. P., Butler P. J. Location and encapsidation of the coat protein cistron of tobacco mosaic virus. A bidirectional elongation of the nucleoprotein rod. Eur J Biochem. 1979 Jan 2;93(1):157–164. doi: 10.1111/j.1432-1033.1979.tb12806.x. [DOI] [PubMed] [Google Scholar]
  27. Ono T., Inoue H., Okada Y. Assembly of rod-shaped virus in vitro: reconstitution with cucumber green mottle mosaic virus protein and tobacco mosaic virus RNA. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3680–3683. doi: 10.1073/pnas.69.12.3680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Perham R. N., Wilson T. M. The characterization of intermediates formed during the disassembly of tobacco mosaic virus at alkaline pH. Virology. 1978 Feb;84(2):293–302. doi: 10.1016/0042-6822(78)90249-0. [DOI] [PubMed] [Google Scholar]
  29. Perham R. N., Wilson T. M. The polarity of stripping of coat protein subunits from the RNA in tobacco mosaic virus under alkaline conditions. FEBS Lett. 1976 Feb 1;62(1):11–15. doi: 10.1016/0014-5793(76)80005-1. [DOI] [PubMed] [Google Scholar]
  30. Richards K. E., Williams R. C. Assembly of tobacco mosaic virus in vitro: effect of state of polymerization of the protein component. Proc Natl Acad Sci U S A. 1972 May;69(5):1121–1124. doi: 10.1073/pnas.69.5.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Richards K. E., Williams R. C. Assembly of tobacco mosaic virus rods in vitro. Elongation of partially assembled rods. Biochemistry. 1973 Nov 6;12(23):4574–4581. doi: 10.1021/bi00747a005. [DOI] [PubMed] [Google Scholar]
  32. Shire S. J., Steckert J. J., Adams M. L., Schuster T. M. Kinetics and mechanism of tobacco mosaic virus assembly: direct measurement of relative rates of incorporation of 4S and 20S protein. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2745–2749. doi: 10.1073/pnas.76.6.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stubbs G., Warren S., Holmes K. Structure of RNA and RNA binding site in tobacco mosaic virus from 4-A map calculated from X-ray fibre diagrams. Nature. 1977 May 19;267(5608):216–221. doi: 10.1038/267216a0. [DOI] [PubMed] [Google Scholar]
  34. WATSON J. D. The structure of tobacco mosaic virus. I. X-ray evidence of a helical arrangement of sub-units around the longitudinal axis. Biochim Biophys Acta. 1954 Jan;13(1):10–19. doi: 10.1016/0006-3002(54)90265-6. [DOI] [PubMed] [Google Scholar]
  35. Zimmern D., Butler P. J. The isolation of tobacco mosaic virus RNA fragments containing the origin for viral assembly. Cell. 1977 Jul;11(3):455–462. doi: 10.1016/0092-8674(77)90064-2. [DOI] [PubMed] [Google Scholar]
  36. Zimmern D. The nucleotide sequence at the origin for assembly on tobacco mosaic virus RNA. Cell. 1977 Jul;11(3):463–482. doi: 10.1016/0092-8674(77)90065-4. [DOI] [PubMed] [Google Scholar]
  37. Zimmern D., Wilson T. M. Location of the origin for viral reassembly on tobacco mosaic virus RNA and its relation to stable fragment. FEBS Lett. 1976 Dec 1;71(2):294–298. doi: 10.1016/0014-5793(76)80954-4. [DOI] [PubMed] [Google Scholar]

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