Summary
The polypeptide composition of extracts of chloroplasts from tobacco leaves systemically infected with different strains of Tobacco Mosaic Virus (TMV) was analyzed by one- and two-dimensional gel electrophoresis. There were no changes in the protein profiles of chloroplasts from infected leaves when compared to control leaves except for the presence of coat protein (CP) of TMV, identified by immunoblotting. When protease-treated intact chloroplasts isolated on Percoll gradients were osmotically disrupted the CP could be detected in both stroma and membrane fractions. The majority of the CP associated with the thylakoid membranes (about 1–5% of the total thylakoid proteins) was in the form of free molecules while stroma contained aggregated or assembled CP (about 0.1% of the soluble proteins). Thylakoid-associated CP was insensitive to protease digestion unless the membranes were first treated with a detergent, indicating that the CP was embedded inside or otherwise complexed with the thylakoid membranes.
Chloroplasts isolated from leaves infected with TMV-PV42, a symptomless strain, contained approximately 10–50 times less CP than did chloroplasts isolated from leaves bearing mosaic symptoms induced by other strains of TMV (U1, PV230 or PV39). A possible role of CP in symptom development is discussed.
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References
Arnon DI: Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15, 1949.
Asselin A, Tremblay C, Bellemare G: Comparative study of the U1, U4 and U7 strains of Tobacco Mosaic Virus. Can J Bot 62:2654–2659, 1984.
Bradford MM: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976.
Bruening G, Beachy RN, Scalla R, Zaitlin M: In vitro and in vivo translation of the ribonucleic acids of a cowpea strain of Tobacco Mosaic Virus. Virology 71:498–517, 1976.
Buchanan BB, Hutcheson SW, Magyarosy AC, Montalbini P: Photosynthesis in healthy and diseased plants. In: Ayres PG (ed) Effects of disease on the physiology of the growing plant. Cambridge University Press, Cambridge, 1981, pp 13–28.
Carroll TW, Kosuge T: Changes in structure of chloroplasts accompanying necrosis of tobacco leaves systemically infected with Tobacco Mosaic Virus. Phytophatol 59:953–962, 1969.
Cline K, Werner-Washburne M, Andrews J, Keegstra K: Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts. Plant Physiol 75:675–678, 1984.
Collmer CW, Vogt VM, Zaitlin M: H protein, a minor protein of TMV virions, contains sequences of the viral coat protein. Virology 126:426–448, 1983.
Ellis RJ: Chloroplast protein synthesis: principle and problems. In: Roodyn D (ed) Subcellular Biochemistry, Vol. 9, Plenum Press, New York, 1983, pp 237–261.
Esau K, Crowshaw J: Relation of Tobacco Mosaic Virus to the host cells. J Cell Biol 33:665–678, 1967.
Finley D, Varsharsky A: The ubiquitin system: functions and mechanisms. T1BS 10:343–347, 1985.
Fish LE, Jagendorf AT: High rates of protein synthesis by isolated chloroplasts. Plant Physiol 70:1107–1114, 1982.
Folk JE: Transglutaminases. Ann Rev Biochem 49:517–531, 1980.
Fraser RRS: Effects of two TMV strains on the synthesis and stability of chloroplast ribosomal RNA in tobacco leaves. Mol Gen Genet 106:73–79, 1969.
Hatta T, Matthews REF: Sites of coat protein accumulation in Turnip Yellow Mosaic Virus-infected cells. Virology 73:1–16, 1976.
Hirai A, Wildman SG: Effect of TMV multiplication on RNA and protein synthesis in tobacco chloroplasts. Virology 38:73–82, 1969.
Holmes FO: A masked strain of Tobacco Mosaic Virus. Phytopathol 24:845–873 1934.
Hoober JK: Chloroplasts. 1st edition Plenum Press, New York, 1984.
Hutcheson SW, Buchanan BB: Bioenergetic and metabolic disturbances in diseased plants. In: Callow JA (ed) Biochemical Plant Pathology. John Wiley & Sons Ltd., New York, 1983, pp 327–345.
Jockush H, Jockush B: Early cell death caused by mutants with defective coat proteins. Mol Gen Genet 102:204–209, 1968.
Johnson EM: Ky. Agric. Exp. Sta. Bull. No. 306, pp 296–297, 1930.
Knight CA: The nature of some of the chemical differences among strains of Tobacco Mosaic Virus J Biol Chem 171:297–308, 1947.
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970.
Lilley RMC, Fitzgerald MP, Rienits KC, Walker DA: Criteria of intactness and the photosynthetic activity of spinach chloroplast preparation. New Phytol 75:1–10, 1975.
Matthews REF: Host-plant responses to virus infection. In: Fraenkel-Conrat H, Wagner RR (eds) Comprehensive Virology, Vol. 16, Plenum Press, New York, 1980, pp 297–359.
Merril CR, Goldman D, VanKeuren ML: Simplified silver protein detection and image enhancement methods in polyacrylamide gels. Electrophoresis 3:17–23, 1982.
Misaghi IJ: Physiology and Biochemistry of Plant-Pathogen Interaction. 1st edition, Plenum Press, New York, 1982.
Nishiguchi M, Nozu Y, Oshima N: Comparative study on coat proteins of an attenuated, a temperature sensitive and their parent strains of Tobacco Mosaic Virus. Phytopath Z 109:104–114, 1984.
Roberts PL, Wood KR: Comparison of protein synthesis in tobacco systemically infected with a severe or a mild strain of Cucumber Mosaic Virus. Phytopath Z 102:257–265, 1981.
Rochon D, Siegel A: Chloroplast DNA transcripts are encapsidated by Tobacco Mosaic Virus coat protein. Proc Natl Acad Sci USA 81:1719–1723, 1984.
Roscoe TJ, Ellis RJ: Two-dimensional gel electrophoresis of chloroplast proteins. In: Edelman M et al. (eds) Methods in Chloroplast Molecular Biology, Elsevier Biomedical Press, Amsterdam, 1982, pp 1015–1028.
Shalla TA, Petersen LJ, Guinchedi L: Partial characterization of virus-like particles in chloroplasts of plants infected with the U5 strain of TMV. Virology 66:94–105, 1975.
Siegel A: Pseudovirions of Tobacco Mosaic Virus. Virology 46:50–59, 1971.
Steinback KE, Mullet JE, Arntzen CJ: Fractionation of thylakoid membrane protein complexes by sucrose densitygradient centrifugation. In: Edelman M et al. (eds) Methods in Chloroplast Molecular Biology. Elsevier Biomedical Press, Amsterdam, 1982, pp 863–872.
Symington J: Electrophoretic transfer of protein from two-dimensional gels to sheets and their detection. In: Bravo R, Celis JE (eds) Two-dimensional gel electrophoresis of proteins. Academic Press, Orlando, 1984, pp 127–168.
Takahashi WN: Increasing the sensitivity of the local lesion method of virus assay. Phytopathol 46:654–656, 1956.
VanRegenmortel MHV: Serology and immunochemistry of plant viruses. Academic Press, New York, 1982.
VanTelgen JH, Van DerZaal EJ, VanLoon LC: Evidence for an association between viral coat protein and host chromatin in mosaic-diseased tobacco leaves. Physiol Plant Pathol 26:83–98, 1985.
Zaitlin M, Israel H: Tobacco Mosaic Virus (type strain). Description of Plant Viruses, no. 151. Commonwealth Mycological Institute, Richmond, Surrey, England and Association of Applied Biologists, Wellesbourne, Warnick, England.
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Reinero, A., Beachy, R.N. Association of TMV coat protein with chloroplast membranes in virus-infected leaves. Plant Mol Biol 6, 291–301 (1986). https://doi.org/10.1007/BF00034936
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DOI: https://doi.org/10.1007/BF00034936