Interaction of tobamovirus movement proteins with the plant cytoskeleton

Science

Volumn 270, Issue 5244, 1995, Pages 1983-1985

  Heinlein, Manfred ^a   Epel, Bernard L ^a,b   Padgett, Hal S ^a,c   Beachy, Roger N ^a  

^a SCRIPPS RESEARCH INSTITUTE   (United States)

^b TEL AVIV UNIVERSITY   (Israel)

^c WASHINGTON UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

HYBRID PROTEIN; VIRUS PROTEIN;

CYTOSKELETON; FLUORESCENCE MICROSCOPY; MICROTUBULE; NONHUMAN; PLANT; PLANT LEAF; PRIORITY JOURNAL; PROTOPLAST; SHORT SURVEY; TOBACCO MOSAIC VIRUS; VIRUS INFECTION; VIRUS INFECTIVITY; VIRUS TRANSMISSION;

NICOTIANA; NICOTIANA SPP.; NICOTIANA TABACUM; TOBACCO MOSAIC VIRUS; TOBAMOVIRUS;

EID: 0029549778     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.270.5244.1983     Document Type: Short Survey

References (54)

1. A. J. Maule, Crit. Rev. Plant Sci. 9, 457 (1991); W. J. Lucas and R. I. Gilbertson, Annu. Rev. Phytopathol. 32, 387 (1994); B. Epel, Plant Mol. Biol. 26, 1343 (1994).
2. A. J. Maule, Crit. Rev. Plant Sci. 9, 457 (1991); W. J. Lucas and R. I. Gilbertson, Annu. Rev. Phytopathol. 32, 387 (1994); B. Epel, Plant Mol. Biol. 26, 1343 (1994).
3. A. J. Maule, Crit. Rev. Plant Sci. 9, 457 (1991); W. J. Lucas and R. I. Gilbertson, Annu. Rev. Phytopathol. 32, 387 (1994); B. Epel, Plant Mol. Biol. 26, 1343 (1994).
4. C. M. Deom, M. J. Oliver, R. N. Beachy, Science 237, 389 (1987).
5. M. C. Deom, M. Lapidot, R. N. Beachy, Cell 69, 221 (1992).
6. K. Tomenius, D. Clapham, T. Meshi, Virology 160, 363 (1987); P. Moore, C. A. Frenczik, C. M. Deom, R. N. Beachy, Protoplasma 170, 115 (1992).
7. K. Tomenius, D. Clapham, T. Meshi, Virology 160, 363 (1987); P. Moore, C. A. Frenczik, C. M. Deom, R. N. Beachy, Protoplasma 170, 115 (1992).
8. S. Wolf, C. M. Deom, R. N. Beachy, W. J. Lucas, Science 246, 377 (1989).
9. C. M. Deom et al., Proc. Natl. Acad. Sci. U.S.A. 87, 3284 (1990).
10. H. S. Padgett and R. N. Beachy, Plant Cell 5, 577 (1993).
11. M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, D. C. Prasher, Science 263, 802 (1994); T. Stearns, Curr. Biol. 5, 262 (1995).
12. M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, D. C. Prasher, Science 263, 802 (1994); T. Stearns, Curr. Biol. 5, 262 (1995).
13. Preparation of protoplasts and electroporation transfer of in vitro-transcribed viral RNA was performed as described [Y. Watanabe, T. Ohno, Y. Okada, Virology 120, 478 (1982); Y. Watanabe, T. Meshi, Y. Okada, FEBS Lett. 219, 65 (1987)]. Infected protoplasts were cultured at 23°C.
14. Preparation of protoplasts and electroporation transfer of in vitro-transcribed viral RNA was performed as described [Y. Watanabe, T. Ohno, Y. Okada, Virology 120, 478 (1982); Y. Watanabe, T. Meshi, Y. Okada, FEBS Lett. 219, 65 (1987)]. Infected protoplasts were cultured at 23°C.
15. Constructs pOb-M:Gfus and pObΔC-GFP were described in B. L. Epel, H. S. Padgett, M. Heinlein, R. Beachy, Gene, in press. We constructed pTMV-M: Gfus by inserting a polymerase chain reaction-amplified portion of the TMV genome containing the MP gene upstream of the GFP gene as an in-frame fusion. Thirty-one nucleotides from the 3′ terminus of the MP open reading frame encoding the coat protein (CP) gene subgenomic promoter were deleted and replaced by 40 nucleotides of the pGEX-KG cloning vector that encode a flexible linker peptide. The gene fusion was inserted into pU3/12-RO-PL, a derivative of the infectious cDNA clone pU3/12 [C. A. Holt and R. N. Beachy, Virology 181, 109 (1991)] encoding wild-type TMV, from which the MP and CP gene sequences had been deleted and replaced with a polycloning site. The resulting construct, pTMV-M:Gfus, contained a fusion of the MP and GFP open reading frames, the transcription of which remained under the control of the subgenomic promoter that controls production of the MP mRNA. The construct pTMVΔC-GFP was designed to express the GFP from the highly active CP promoter and was constructed by subcloning the GFP gene from pGFP-01 into the Sna BI site of pTMVΔC-SB, a derivative of pU3/12 from which CP gene sequences have been deleted (M. Heinlein, B. L. Epel, H. S. Padgett, R. N. Beachy, unpublished results).
16. Constructs pOb-M:Gfus and pObΔC-GFP were described in B. L. Epel, H. S. Padgett, M. Heinlein, R. Beachy, Gene, in press. We constructed pTMV-M: Gfus by inserting a polymerase chain reaction-amplified portion of the TMV genome containing the MP gene upstream of the GFP gene as an in-frame fusion. Thirty-one nucleotides from the 3′ terminus of the MP open reading frame encoding the coat protein (CP) gene subgenomic promoter were deleted and replaced by 40 nucleotides of the pGEX-KG cloning vector that encode a flexible linker peptide. The gene fusion was inserted into pU3/12-RO-PL, a derivative of the infectious cDNA clone pU3/12 [C. A. Holt and R. N. Beachy, Virology 181, 109 (1991)] encoding wild-type TMV, from which the MP and CP gene sequences had been deleted and replaced with a polycloning site. The resulting construct, pTMV-M:Gfus, contained a fusion of the MP and GFP open reading frames, the transcription of which remained under the control of the subgenomic promoter that controls production of the MP mRNA. The construct pTMVΔC-GFP was designed to express the GFP from the highly active CP promoter and was constructed by subcloning the GFP gene from pGFP-01 into the Sna BI site of pTMVΔC-SB, a derivative of pU3/12 from which CP gene sequences have been deleted (M. Heinlein, B. L. Epel, H. S. Padgett, R. N. Beachy, unpublished results).
17. Constructs pOb-M:Gfus and pObΔC-GFP were described in B. L. Epel, H. S. Padgett, M. Heinlein, R. Beachy, Gene, in press. We constructed pTMV-M: Gfus by inserting a polymerase chain reaction-amplified portion of the TMV genome containing the MP gene upstream of the GFP gene as an in-frame fusion. Thirty-one nucleotides from the 3′ terminus of the MP open reading frame encoding the coat protein (CP) gene subgenomic promoter were deleted and replaced by 40 nucleotides of the pGEX-KG cloning vector that encode a flexible linker peptide. The gene fusion was inserted into pU3/12-RO-PL, a derivative of the infectious cDNA clone pU3/12 [C. A. Holt and R. N. Beachy, Virology 181, 109 (1991)] encoding wild-type TMV, from which the MP and CP gene sequences had been deleted and replaced with a polycloning site. The resulting construct, pTMV-M:Gfus, contained a fusion of the MP and GFP open reading frames, the transcription of which remained under the control of the subgenomic promoter that controls production of the MP mRNA. The construct pTMVΔC-GFP was designed to express the GFP from the highly active CP promoter and was constructed by subcloning the GFP gene from pGFP-01 into the Sna BI site of pTMVΔC-SB, a derivative of pU3/12 from which CP gene sequences have been deleted (M. Heinlein, B. L. Epel, H. S. Padgett, R. N. Beachy, unpublished results).
18. Microscopy was performed with a Nikon Optiphot 2UD microscope. For visualization of GFP fluorescence, a FITC filter cube (470- to 490-nm excitation filter, 505-nm dichroic mirror, and 520-nm barrier filter) was used.
19. For immunostaining, protoplasts were harvested at 16 hpi, fixed for 30 min in 50 mM phosphate buffer (pH 6.7) containing 3% paraformaldehyde and 5 mM EGTA, and then spun onto polylysine-coated slides and dried. The protoplasts were then extracted for 10 min with cold methanol. Antibodies were applied in phopshate-buffered saline, pH 7.0, containing 0.5% Tween-20 and 5 mM EGTA, and all washes were performed in the same buffer. The samples were mounted in Mowiol (Calbiochem) containing 2.5% 1,4-diazobicyclo-[2.2.2.]-octane (DABCO) as an antifade reagent.
20. A 510-to 560-nm excitation filter, a 575-nm dichroic mirror, and a 590-nm barrier filter were used to examine rhodamine fluorescence.
21. M. Heinlein, B. L. Epel, H. S. Padgett, R. N. Beachy, data not shown.
22. C. W. Lloyd, A. R. Slabas, A. J. Powell, S. B. Lowe, Planta 147, 500 (1980); R. E. Williamson, Int. Rev. Cytol. 129, 135 (1991).
23. C. W. Lloyd, A. R. Slabas, A. J. Powell, S. B. Lowe, Planta 147, 500 (1980); R. E. Williamson, Int. Rev. Cytol. 129, 135 (1991).
24. T. Akashi, S. Kawasaki, H. Shibaoka, Planta 182, 363 (1990).
25. Prefixation of F-actin and buffer conditions were as described [J. Katsuta, Y. Hashiguchi, H. Shibaoka, J. Cell Sci. 95, 413 (1990)].
26. P. K. Hepler, Protoplasma 111, 121 (1982); K. J. Oparka, D. A. M. Prior, J. W. Crawford, Plant Cell Env. 17, 163 (1994).
27. P. K. Hepler, Protoplasma 111, 121 (1982); K. J. Oparka, D. A. M. Prior, J. W. Crawford, Plant Cell Env. 17, 163 (1994).
28. T. Akashi and H. Shibaoka, J. Cell Sci. 98, 169 (1991); H. Quader, A. Hoffman, E. Schnepf, Eur. J. Cell Biol. 44, 17 (1987); I. K. Lichtscheidl and W. G. Url, Protoplasma 157, 203 (1990); I. K. Lichtscheidl, S. A. Lancelle, P. K. Hepler, ibid. 155, 116 (1990).
29. T. Akashi and H. Shibaoka, J. Cell Sci. 98, 169 (1991); H. Quader, A. Hoffman, E. Schnepf, Eur. J. Cell Biol. 44, 17 (1987); I. K. Lichtscheidl and W. G. Url, Protoplasma 157, 203 (1990); I. K. Lichtscheidl, S. A. Lancelle, P. K. Hepler, ibid. 155, 116 (1990).
30. T. Akashi and H. Shibaoka, J. Cell Sci. 98, 169 (1991); H. Quader, A. Hoffman, E. Schnepf, Eur. J. Cell Biol. 44, 17 (1987); I. K. Lichtscheidl and W. G. Url, Protoplasma 157, 203 (1990); I. K. Lichtscheidl, S. A. Lancelle, P. K. Hepler, ibid. 155, 116 (1990).
31. T. Akashi and H. Shibaoka, J. Cell Sci. 98, 169 (1991); H. Quader, A. Hoffman, E. Schnepf, Eur. J. Cell Biol. 44, 17 (1987); I. K. Lichtscheidl and W. G. Url, Protoplasma 157, 203 (1990); I. K. Lichtscheidl, S. A. Lancelle, P. K. Hepler, ibid. 155, 116 (1990).
32. P. K. Hepler, B. A. Palevitz, S. A. Lancelle, M. M. McCauley, I. Lichtscheidl, J. Cell Sci. 96, 355 (1990).
33. T. A. Schroer and M. P. Sheetz, Annu. Rev. Physiol. 53, 629 (1990); J. Lippincott-Schwartz, N. B. Cole, A. Marotta, P. A. Conrad, G. S. Bloom, J. Cell. Biol. 128, 293 (1995).
34. T. A. Schroer and M. P. Sheetz, Annu. Rev. Physiol. 53, 629 (1990); J. Lippincott-Schwartz, N. B. Cole, A. Marotta, P. A. Conrad, G. S. Bloom, J. Cell. Biol. 128, 293 (1995).
35. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
36. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
37. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
38. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
39. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
40. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
41. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
42. K. Kristensson, E. Lycke, M. Ryotta, B. Svennerholm, A. Vahlne, J. Gen. Virol. 67, 2023 (1986); S. Dales and Y. Chardonnet, Virology 56, 465 (1973); A. P. de Matos and Z. G. Carvalho, Biol. Cell 78, 229 (1993); U. I. Heine, A. E. Demsey, R. W. Tucker, A. F. Bykovsky, J. Gen. Virol. 66, 275 (1985); E. Lycke and H. Tsiang, J. Virol. 61, 273 (1987); K. S. Topp, L. B. Meade, J. H. LaVail, J. Neurosci. 14, 318 (1994); B. Norrild, V. P. Lehto, I. Virtanen, J. Gen Virol. 67, 97 (1986); M. J. Rindler, I. E. Ivanov, D. D. Sabatini, J. Cell Biol. 104, 231 (1987).
43. A. V. Karasev, A. S. Kashina, V. I. Gelfand, V. V. Dolja, FEBS Lett. 304, 12 (1992).
44. M. Terasaki, L. B. Chen, K. Fujiwara, J. Cell Biol. 103, 1557 (1986); S. L. Dabora and M. P. Scheetz, Cell 54, 27 (1988); R. D. Vale and H. Hotani, J. Cell Biol. 107, 2233 (1988).
45. M. Terasaki, L. B. Chen, K. Fujiwara, J. Cell Biol. 103, 1557 (1986); S. L. Dabora and M. P. Scheetz, Cell 54, 27 (1988); R. D. Vale and H. Hotani, J. Cell Biol. 107, 2233 (1988).
46. M. Terasaki, L. B. Chen, K. Fujiwara, J. Cell Biol. 103, 1557 (1986); S. L. Dabora and M. P. Scheetz, Cell 54, 27 (1988); R. D. Vale and H. Hotani, J. Cell Biol. 107, 2233 (1988).
47. S. A. Lancelle, M. Cresti, P. K. Hepler, Protoplasma 140, 141 (1987).
48. W. Knebel, H. Quader, E. Schnepf, Eur. J. Cell Biol. 52, 328 (1990).
49. V. Citovsky et al., Cell 60, 637 (1990); V. Citovsky et al., Plant Cell 4, 397 (1992).
50. V. Citovsky et al., Cell 60, 637 (1990); V. Citovsky et al., Plant Cell 4, 397 (1992).
51. W. E. Theurkauf, B. M. Alberts, Y. N. Jan, T. A. Jongens, Development 118, 1169 (1993); J. E. Wilhelm and R. D. Vale, J. Cell Biol. 123, 269 (1993).
52. W. E. Theurkauf, B. M. Alberts, Y. N. Jan, T. A. Jongens, Development 118, 1169 (1993); J. E. Wilhelm and R. D. Vale, J. Cell Biol. 123, 269 (1993).
53. K. Ainger et al., J. Cell Biol. 123, 431 (1993).
54. We thank J. Harper for providing pGFP-01, which contains the GFP coding sequence obtained from R. Tsien, and G. Klier for assistance with confocal microscopy. Supported by NSF grants MCB 9209530 and MCB 9317368 and the Scripps Family Chair to R.N.B. M.H. was supported by the Deutsche Forschungsgemeinschaft and the Human Frontier Science Program Organization.