Summary
Infection ofNicotiana clevelandii protoplasts by raspberry ringspot nepovirus resulted in the accumulation of about 24 polypeptides that differed in Mr and pI from polypeptides accumulating in mock-inoculated protoplasts. Similar polypeptides accumulated in protoplasts infected with the S and E strains of RRV but different infection-specific polypeptides were detected in protoplasts infected with tobacco ringspot nepovirus. The Mr of RRV-specific polypeptides ranged from 210 000 to 18 000 and most are presumed to be derived from others by proteolytic cleavage. No evidence was found for marked changes in polypeptide abundance with time after inoculation or for any virus-specific polypeptide becoming disproportionately abundant in the medium during culture.
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References
Acosta O, Mayo MA (1990) Accumulation of different types of raspberry ringspot nepovirus particle in infectedNicotiana protoplasts. J Gen Virol 71: 713–717
Acosta O, Mayo MA (1990) Unusual electrophoretic properties of the coat protein of raspberry ringspot nepovirus. Intervirology 31: 31–37
Ames GFL, Nikaido K (1976) Two-dimensional gel electrophoresis of membrane proteins. Biochemistry 15: 616–623
Barker H, Harrison BD (1977) Infection of tobacco mesophyll protoplasts with raspbery ringspot virus alone and together with tobacco rattle virus. J Gen Virol 35: 125–133
Blok VC, Wardell J, Jolly CA, Manoukian A, Robinson DJ, Edwards ML, Mayo MA (1992) The nucleotide sequence of RNA-2 of raspberry ringspot nepovirus. J Gen Virol 73: 2189–2194
Blum H, Beier H, Gross HJ (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8: 93–99
Colas des Francs C, Thiellement H, De Vienne D (1985) Analysis of leaf proteins by two-dimensional gel electrophoresis. Protease action as exemplified by ribulose bisphosphate carboxylase/oxygenase degradation and procedure to avoid proteolysis during extraction. Plant Physiol 78: 178–182
Damerval C, De Vienne D, Zivy M, Thiellement H (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat seedling proteins. Electrophoresis 7: 52–54
Demangeat G, Greif C, Hemmer O, Fritsch C (1990) Analysis of the in vitro cleavage products of the tomato black ring virus RNA-1-encoded 250 K polyprotein. J Gen Virol 71: 1649–1654
Demangeat G, Hemmer O, Fritsch C, Le Gall O, Candresse T (1991) In vitro processing of the RNA-2-encoded polyprotein of two nepoviruses: tomato black ring virus and grapevine chrome mosaic virus. J Gen Virol 72: 247–252
Demangeat G, Hemmer O, Reinbolt J, Mayo MA, Fritsch C (1992) Virus-specific proteins in cells infected with tomato black ring nepovirus: evidence for proteolytic processing in vivo. J Gen Virol 73: 1609–1614
Dunbar BD (1987) Two-dimensional electrophoresis and immunological methods. Plenum, New York
Duncan R, McConkey EH (1982) How many proteins are there in a typical mammalian cell? Clin Chem 28: 749–755
Fritsch C, Mayo MA, Murant AF (1980) Translation products of genome and satellite RNAs of tomato black ring virus. J Gen Virol 46: 381–389
Goldbach R, Rezelman G (1983) Orientation of the cleavage map of the 200-kilodalton polypeptide encoded by the bottom-component RNA of cowpea mosaic virus. J Virol 46: 614–619
Greif C, Hemmer O, Fritsch C (1988) Nucleotide sequence of tomato black ring virus RNA-1. J Gen Virol 69: 1517–1529
Harrison BD, Mayo MA (1983) The use of protoplasts in plant virus research. In: Helgeson JP, Deveral BJ (eds) Use of tissue culture and protoplasts in plant pathology. Academic Press, London, pp 69–137
Harrison BD, Murant AF, Mayo MA (1972) Two properties of raspberry ringspot virus determined by its smaller RNA. J Gen Virol 17: 137–141
Harrison BD, Murant AF, Mayo MA, Roberts IM (1974) Distribution of determinants for symptom production, host range and nematode transmissibility between the two RNA components of raspberry ringspot virus. J Gen Virol 22: 233–247
Jones AT, Mayo MA, Henderson SJ (1985) Biological and biochemical properties of an isolate of cherry rasp leaf virus from red raspberry. Ann Appl Biol 106: 101–110
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Mayo MA, Barker H, Harrison BD (1982) Specificity and properties of the genomelinked proteins of nepoviruses. J Gen Virol 59: 149–162
Meyer M, Hemmer O, Mayo MA, Fritsch C (1986) The nucleotide sequence of tomato black ring virus RNA-2. J Gen Virol 67: 1257–1271
Murant AF, Taylor M, Duncan G, Rashke JH (1981) Improved estimates of molecular weight of plant virus RNA by agarose gel electrophoresis and electron microscopy. J Gen Virol 53: 321–332
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021
Robinson DJ, Mayo MA (1977) Changing rates of uptake of [3H]leucine and other compounds during culture of tobacco mesophyll protoplasts. Plant Sci Lett 8: 197–204
Robinson DJ, Barker H, Harrison BD, Mayo MA (1980) Replication of RNA-1 of tomato black ring virus independently of RNA-2. J Gen Virol 51: 317–326
Vlak JM, Smith GE, Summers MD (1981) Selection and in vitro translation ofAutographa californica nuclear polyhedrosis virus mRNA. J Virol 40: 762–771
Wellink J, Jaegle M, Goldbach R (1987) Detection of a novel protein encoded by the bottom-component RNA of cowpea mosaic virus, using antibodies raised against a synthethic peptide. J Virol 61: 236–238
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Acosta, O., Mayo, M.A. Polypeptide synthesis induced inNicotiana clevelandii protoplasts by infection with raspberry ringspot nepovirus. Archives of Virology 129, 145–154 (1993). https://doi.org/10.1007/BF01316891
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DOI: https://doi.org/10.1007/BF01316891