1887

Journal of General Virology

Volume 82, Issue 6

Visualization by atomic force microscopy of tobacco mosaic virus movement protein–RNA complexes formed Free

Abstract

The structure of complexes formed by tobacco mosaic virus (TMV)-coded movement protein (MP) with TMV RNA and short (890 nt) synthetic RNA transcripts was visualized by atomic force microscopy on a mica surface. MP molecules were found to be distributed along the chain of RNA and the structure of MP–RNA complexes depended on the molar MP:RNA ratios at which the complexes were formed. A rise in the molar MP:TMV RNA ratio from 20:1 to 60–100:1 resulted in an increase in the density of the MP packaging on TMV RNA and structural conversion of complexes from RNase-sensitive ‘beads-on-a-string’ into a ‘thick string’ form that was partly resistant to RNase. The ‘thick string’-type RNase-resistant complexes were also produced by short synthetic RNA transcripts at different MP:RNA ratios. The ‘thick string’ complexes are suggested to represent clusters of MP molecules cooperatively bound to discrete regions of TMV RNA and separated by protein-free RNA segments.

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© Microbiology Society
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2001-06-01
2019-10-14

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References

  1. Binnig, G., Quate, C. F. & Gerber, C. ( 1986; ). Atomic force microscope. Physical Review Letters 56, 930-933.[CrossRef]
    [Google Scholar]
  2. Citovsky, V. ( 1999; ). Tobacco mosaic virus: a pioneer of cell-to-cell movement. Philosophical Transactions of the Royal Society of London B Biological Sciences 354, 637-647.[CrossRef]
    [Google Scholar]
  3. Citovsky, V., Knorr, D., Schuster, G. & Zambryski, P. ( 1990; ). The P30 movement protein of tobacco mosaic virus is a single strand nucleic acid binding protein. Cell 60, 637-647.[CrossRef]
    [Google Scholar]
  4. Citovsky, V., Wong, M., Show, A., Prasad, B. V. & Zambryski, P. ( 1992; ). Visualization and characterization of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell 4, 397-411.[CrossRef]
    [Google Scholar]
  5. Drygin, Yu. F., Bordunova, O. A., Gallyamov, M. O. & Yaminsky, I. V. ( 1998; ). Atomic force microscopy examination of TMV and virion RNA. FEBS Letters 425, 217-221.[CrossRef]
    [Google Scholar]
  6. Filonov, A. S. & Yaminsky, I. V. (1997). SPM Control and Image Processing Software. Chichester: Advanced Technologies Center, Moscow.
  7. Ghoshroy, S., Lartey, R., Sheng, J. & Citovsky, V. ( 1997; ). Transport of proteins and nucleic acids through plasmodesmata. Annual Review of Plant Physiology and Plant Molecular Biology 48, 27-50.[CrossRef]
    [Google Scholar]
  8. Heinlein, M., Epel, B. L., Padgett, H. S. & Beachy, R. N. ( 1995; ). Interaction of tobamovirus movement proteins with the plant cytoskeleton. Science 270, 1983-1985.[CrossRef]
    [Google Scholar]
  9. Heinlein, M., Padgett, H. S., Gens, J. S., Pickard, B. G., Casper, S. J., Epel, B. L. & Beachy, R. N. ( 1998; ). Changing patterns of localization of tobacco mosaic movement protein and replicase to the endoplasmic reticulum and microtubules during infection. Plant Cell 10, 1107-1120.[CrossRef]
    [Google Scholar]
  10. Jafri, S., Evoy, S., Cho, K., Craighead, H. G. & Winans, S. C. ( 1999; ). An Lpr-type transcriptional regulator from Agrobacterium tumefaciens condenses more than 100 nucleotides of DNA into globular nucleoprotein complexes. Journal of Molecular Biology 288, 811-824.[CrossRef]
    [Google Scholar]
  11. Karpova, O. V., Ivanov, K. I., Rodionova, N. P., Dorokhov, Yu. L. & Atabekov, J. G. ( 1997; ). Nontranslatability and dissimilar behavior in plants and protoplasts of viral RNA and movement protein complexes formed in vitro. Virology 230, 11-21.[CrossRef]
    [Google Scholar]
  12. Karpova, O. V., Rodionova, N. P., Ivanov, K. I., Kozlovsky, S. V., Dorokhov, Yu. L. & Atabekov, J. G. ( 1999; ). Phosphorylation of tobacco mosaic virus movement protein abolishes its translation repressing ability. Virology 261, 20-24.[CrossRef]
    [Google Scholar]
  13. Kiselyova, O. I. & Yaminsky, I. V. ( 1999; ). Proteins and membrane-protein complexes. Colloid Journal 61, 1-19.
     [Google Scholar]
  14. Lazarovitz, V. & Beachy, R. N. ( 1999; ). Viral proteins as probes for intracellular and intercellular tracking in plants. Plant Cell 11, 535-548.[CrossRef]
    [Google Scholar]
  15. Li, Q. & Palukaitis, P. ( 1996; ). Comparison of the nucleic acid- and NTP-binding properties of the movement protein of cucumber mosaic cucumovirus and tobacco mosaic tobamovirus. Virology 216, 71-79.[CrossRef]
    [Google Scholar]
  16. Lucas, W. J. & Gilbertson, R. L. ( 1994; ). Plasmodesmata in relation to viral movement within leaf tissues. Annual Review of Phytopathology 32, 387-411.[CrossRef]
    [Google Scholar]
  17. Lyubchenko, Yu. L., Shlyakhtenko, L. S., Harrington, R. E., Oden, P. I. & Lindsay, S. M. ( 1993; ). Atomic force microscopy of long DNA: imaging in air and under water. Proceedings of the National Academy of Sciences, USA 90, 2137-2140.[CrossRef]
    [Google Scholar]
  18. Lyubchenko, Yu. L., Jacobs, B. L., Lindsay, S. M. & Stasiak, A. ( 1995; ). Atomic force microscopy of nucleoprotein complexes. Scanning Microscopy 9, 705-727.
     [Google Scholar]
  19. McLean, B. G., Zupan, J. & Zambryski, P. ( 1995; ). Tobacco mosaic movement protein associates with the cytoskeleton in tobacco cells. Plant Cell 7, 2101-2114.[CrossRef]
    [Google Scholar]
  20. Shlyakhtenko, L. S., Potaman, V. N., Sinden, R. R. & Lyubchenko, Yu. L. ( 1998; ). Structure and dynamics of supercoil-stabilized DNA cruciforms. Journal of Molecular Biology 280, 61-72.[CrossRef]
    [Google Scholar]
  21. Stemmer, A. & Engel, A. ( 1990; ). Imaging biological macromolecules by STM: quantitative interpretation of topographs. Ultramicroscopy 34, 129-140.[CrossRef]
    [Google Scholar]
  22. Tzfira, T., Rhee, Y., Chen, M.-H., Kunik, T. & Citovsky, V. ( 2000; ). Nucleic acid transport in plant–microbe interactions: the molecules that walk through the walls. Annual Review of Microbiology 54, 187-219.[CrossRef]
    [Google Scholar]
  23. Valle, M., Valpuesta, J. M., Carrascosa, J. L., Tamayo, J. & Garcia, R. ( 1996; ). The interaction of DNA with bacteriophage 29 connector: a study by AFM and TEM. Journal of Structural Biology 116, 390-398.[CrossRef]
    [Google Scholar]
  24. Waigmann, E., Lucas, W., Citovsky, V. & Zambrysky, P. ( 1994; ). Direct functional assay for tobacco mosaic virus cell-to-cell movement protein and identification of a domain involved in increasing plasmodesmal permeability. Proceedings of the National Academy of Sciences, USA 91, 1433-1437.[CrossRef]
    [Google Scholar]
  25. Wolf, S., Deom, C. M., Beachy, R. N. & Lucas, W. Y. ( 1989; ). Movement protein of tobacco mosaic virus modifies plasmodesmal size exclusion limit. Science 246, 377-379.[CrossRef]
    [Google Scholar]
  26. Yodh, J. G., Lyubchenko, Y. L., Shlyakhtenko, L. S., Woodbury, N. & Lohr, D. ( 1999; ). Evidence for nonrandom behavior in 208–12 subsaturated nucleosomal array populations analyzed by AFM. Biochemistry 38, 15756-15763.[CrossRef]
    [Google Scholar]
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Visualization by atomic force microscopy of tobacco mosaic virus movement protein–RNA complexes formed in vitro
J. Gen. Virol. 82, 1503 (2001); https://doi.org/10.1099/0022-1317-82-6-1503
/content/journal/jgv/10.1099/0022-1317-82-6-1503
/content/journal/jgv/10.1099/0022-1317-82-6-1503
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