Tobacco mosaic virus particles

Assembly and Stability of the Tobacco Mosaic Virus Particle D. L. D. Caspar... 

Figure 4.21 Effect of axial ratio and particle concentration on relative viscosity. Data are for tobacco mosaic virus particles. Adapted from M. A. Lauffer, 7. Am. Chem. Soc., 66, 1188. Copyright 1944 by The American Chemical Society, Inc.

The ratio (a/b), called the axial ratio of the ellipsoid, is frequently used as a measure of the deviation from sphericity of a particle. It plays an important role, for example, in our discussions of sedimentation and viscosity in Chapters 2 and 4, respectively. In the event that a > b, the prolate ellipsoid approximates a cylinder and, as such, is often used to describe rod-shaped particles such as the tobacco mosaic virus particles shown in Figure 1.12a. Likewise, if a < b, the oblate ellipsoid approaches the shape of a disk. Thus, even the irregular clay platelets of Figure 1.12b may be approximated as oblate ellipsoids. 

FIG. 1.12 Electron micrograph of two different types of particles that represent extreme variations from spherical particles (a) tobacco mosaic virus particles (Photograph courtesy of Carl Zeiss, Inc., New York) and (b) clay particles (sodium kaolinite) of mean diameter 0.2 fim (by matching circular fields). In both (a) and (b), contrast has been enhanced by shadow casting (see Section 1.6a.2a and Figure 1.21). (Adapted from M. D. Luh and R. A. Bader, J. Colloid Interface Sci. 33, 539(1970). 

Figure 4.12a shows plots of the intrinsic viscosity —in volume fraction units —as a function of axial ratio according to the Simha equation. Figure 4.12b shows some experimental results obtained for tobacco mosaic virus particles. These particles —an electron micrograph of which is shown in Figure 1.12a—can be approximated as prolate ellipsoids. Intrinsic viscosities are given by the slopes of Figure 4.12b, and the parameters on the curves are axial ratios determined by the Simha equation. Thus we see that particle asymmetry can also be quantified from intrinsic viscosity measurements for unsolvated particles. 

S-TT Klug, A., The tobacco mosaic virus particle structure and assembly , Phil. Trans. RoyalSoc. Ser. B, Biol. Scl. 1999, 354, 531-535. 

Stubbs, G. (1999). Tobacco mosaic virus particle structure and the initiation of disassembly. Philos. Trans. R. Soc. Lond. B Biol. Sci. 354, 551-557. 

The Size and Shape of Tobacco Mosaic Virus Particles. J. Amer. chem. 

Figure 1.3.3 Electron micrograph of tobacco mosaic virus particle. [Courtesy of Prof. Emeritus Robley C. Williams, Virus Laboratory and Dept, of Molecular Biology, Univ. of California, Berkeley.]...

Figure 3. Transmission electron microscopy of enzyme-cleaved, tobacco mosaic virus particles on the apex of a tungsten field-emitter tip (imaged at 200kV). TMV sample kindly supplied by P. J. Butler, the MRC, Cambridge, England.

Kuznetsov, Y. G., Larson, S. B., Day, J., Greenwood, A., and McPherson, A. 2001. Structural transitions of satellite tobacco mosaic virus particles. Virology 284,223-234. 

Klug, A. The tobacco mosaic virus particle Structure and... 

Each tobacco mosaic virus particle consists of one long thread of nucleic acid embedded in protein. The protein surrounds the nucleic acid in loops or in the fashion of screw threads making up the super-molecule. Treatment with phenol separates nucleic acids from protein. The nucleic acid obtained in this mild way remains infectious, and in a host cell can cause virus multiplication and consequent symptoms of disease. About 95% of the material is protein it consists of individual subunits with a molecular weight of 17,500, which exhibit a marked tendency to aggregate At neutral or slightly acidic pH the protein molecules aggregiite to little rozls, very similar to the intact virus particles both in shape and size. The amino acid sequence is now known. Mutants obtained by nitrous acid treatment (see above) show up differences in the amino acid sequence usually only one amino acid has been replaced, for example, serine by leucine, or leucine by phenylalanine. 

Some virus particles have their protein subunits symmetrically packed in a helical array, forming hollow cylinders. The tobacco mosaic virus (TMV) is the classic example. X-ray diffraction data and electron micrographs have revealed that 16 subunits per turn of the helix project from a central axial hole that runs the length of the particle. The nucleic acid does not lie in this hole, but is embedded into ridges on the inside of each subunit and describes its own helix from one end of the particle to the other. 

Figure 5.4 Structure and manner of assembly of a simple virus, tobacco mosaic virus, (a) Electron micrograph at high resolution of a portion of the virus particle, (b) Assembly of the tobacco mosaic virion. The RNA assumes a helical configuration surrounded by the protein capsomeres. The center of the particle is hollow.

A typical virus with helical symmetry is the tobacco mosaic virus (TMV). This is an RNA virus in which the 2130 identical protein subunits (each 158 amino acids in length) are arranged in a helix. In TMV, the helix has 16 1/2 subunits per turn and the overall dimensions of the virus particle are 18 X 300 nm. The lengths of helical viruses are determined by the length of the nucleic acid, but the width of the helical virus particle is determined by the size and packing of the protein subunits. 

Completely different mechanisms are involved in the self-assembly of the tobacco mosaic virus (TMV). This virus consists of single-strand RNA, which is surrounded by 2,130 identical protein units, each of which consists of 158 amino acid residues. A virus particle, which requires the tobacco plant as a host, has a rodlike structure with helical symmetry ( Stanley needles ). It is 300 nm long, with a diameter of 18nm. The protein and RNA fractions can be separated, and the viral... 

True self-assembly is observed in the formation of many oligomeric proteins. Indeed, Friedman and Beychok reviewed efforts to define the subunit assembly and reconstitution pathways in multisubunit proteins, and all of the several dozen examples cited in their review represent true self-assembly. Polymeric species are also formed by true self-assembly, and the G-actin to F-actin transition is an excellent example. By contrast, there are strong indications that ribosomal RNA species play a central role in specifying the pathway to and the structure of ribosome particles. And it is interesting to note that the assembly of the tobacco mosaic virus (TMV) appears to be a two-step hybrid mechanism the coat protein subunits first combine to form 34-subunit disks by true self-assembly from monomeric and trimeric com-... 

Another complex macromolecular aggregate that can reassemble from its components is the bacterial ribosome. These ribosomes are composed of 55 different proteins and by 3 different RNA molecules, and if the individual components are incubated under appropriate conditions in a test tube, they spontaneously form the original structure (Alberts et al., 1989). It is also known that even certain viruses, e.g., tobacco mosaic virus, can reassemble from the components this virus consists of a single RNA molecule contained in a protein coat composed by an array of identical protein subunits. Infective virus particles can self-assemble in a test tube from the purified components. 

Figure 7-8 (A) Electron micrograph of the rod-shaped particles of tobacco mosaic virus. Omikron, Photo Researchers. See also Butler and Klug.42 (B) A stereoscopic computer graphics image of a segment of the 300 nm long tobacco mosaic virus. The diameter of the rod is 18 nm, the pitch of the helix is 2.3 nm, and there are 16 1 3 subunits per turn. The coat is formed from 2140 identical 17.5-kDa subunits. The 6395-nucleotide genomic RNA is represented by the dark chain exposed at the top of the segment. The resolution is 0.4 nm. From Namba, Caspar, and Stubbs.47 (C) A MolScript ribbon drawing of two stacked subunits. From Wang and Stubbs.46...

Several colloidal systems containing anisodimensional particles, such as iron oxide and tobacco mosaic virus sols, show reversible... 

We have already dealt with some general aspects of biochemical self-assembly in Section 2.10 including the remarkable formation of viral capsids. There are some biochemical examples, however, that translate readily into supramolecular chemical concepts and have been pivotal in defining the field. One such system is the tobacco mosaic virus, a virus that is very harmful to a variety of crops including tobacco, tomato, pepper, cucumbers and species such as ornamental flowers. This system consists of a helical virus particle measuring some 300 X 18 nm (Figure 10.6). A central strand of RNA is sheathed by 2130 identical protein subunits, each of which contains 158 amino acids. What is remarkable about... 

Certain viruses, notably tobacco mosaic virus (TMV), can also self-assemble. A TMV particle can be dissociated into its component proteins and RNA and then reassembled into infective virus particles on mixing the components together again. 

Regarding the mass range, DNA ions of 108 Da were weighed by mass spectrometry [77], In the same way, non-covalent complexes with molecular weights up to 2.2 MDa were measured by mass spectrometry [78], Intact viral particles of tobacco mosaic virus with a theoretical molecular weight of 40.5 MDa were analysed with an electrospray ionization charge detection time-of-flight mass spectrometer [6]. 

Fig. 12. Mass spectra of Rice Yellow Mottle Virus (RYMV) and Tobacco Mosaic Virus (TMV) particles analyzed with an electrospray ionization charge detection time-of-flight mass spectrometer. Inset, electron micrographs of the icosahedral RYMV (diameter 28.8 nm) and the cylindrical TMV (-300 nm long and 17 nm in diameter). The known molecular weight of RYMV and TMV are 6.5X106 and 40.5X106 Daltons,respectively...

---