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Translation in a reticulocyte cell-free system of RNA isolated from blood and culture forms of Trypanosoma brucei

Published online by Cambridge University Press:  06 April 2009

M. J. Eggitt ,
L. Tappenden  and
K. N. Brown
M. J. Eggitt
Affiliation:
Division of Parasitology, National Institute for Medical Research, Mill Hill, LondonNW7 1AA
L. Tappenden
Affiliation:
Division of Parasitology, National Institute for Medical Research, Mill Hill, LondonNW7 1AA
K. N. Brown
Affiliation:
Division of Parasitology, National Institute for Medical Research, Mill Hill, LondonNW7 1AA
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Extract

RNA with messenger activity has been extracted from both blood and culture (insect mid–gut) forms of Trypanosoma brucei and translated in a reticulocyte cell–free system. The products of this cell–free system have been compared, and many common polypeptides demonstrated. A major polypeptide of 58000–65000 molecular weight was made when both blood and culture from RNA was added to the cell–free system.

Antiserum raised against purified variant antigen from a cloned variant (MIAG 099) was used to detect specific products of this system. A major polypeptide of approximately 58000–65000 molecular weight was precipitated when the homologous trypanosome (MIAG 099) blood form RNA was used in the cell–free system. No such polypeptide was precipitated when RNA from a heterologous strain culture or blood form was used in the system. Competition experiments, in which excess purified variant antigen was added after incubation but before addition of specific antiserum, confirmed that the polypeptide of 58000–65000 molecular weight is the variant antigen.

Poly(A)–containing RNA from trypanosomes was shown to contain the mRNA for the variant antigen.

Type
Research Article
Information
Parasitology , Volume 75 , Issue 2 , October 1977 , pp. 133 - 141
Copyright
Copyright © Cambridge University Press 1977

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References

Allsopp, B. A., Njogu, A. R. & Humpkryes, K. C. (1971). Nature and location of Trypanosoma brucei subgroup exoantigen and its relationship to 4s antigen. Experimental Parasitology 29, 271–84.CrossRefGoogle ScholarPubMed
Berridge, M. V., Farmer, S. R., Green, CD., Henshaw, E. C. & Tata, J. R. (1976). Characterization of polysomes from Xenopus liver synthesizing vitellogenin and translation of vitellogenin and albumin messenger RNA's in vitro. European Journal of Biochemistry 62, 161–71.CrossRefGoogle ScholarPubMed
Bridgen, P. J., Cross, G. A. M. & Bridgen, J. (1976). N–terminal amino acid sequences of variant specific antigens from Trypanosoma brucei. Nature, London 263, 613–14.CrossRefGoogle ScholarPubMed
Brown, R. C, Evans, D. A. & Vickerman, K. (1973). Changes in oxidative metabolism and ultrastructure accompanying differentiation of the mitochondrion. in Trypanosoma brucei. International Journal for Parasitology 3, 691–704.CrossRefGoogle ScholarPubMed
Campbell, P. N. & Blobel, G. (1976). The role of organelles in the chemical modification of the primary translation products of secretory proteins. FEBS Letters 72, 215–26.CrossRefGoogle ScholarPubMed
Chesters, J. K. (1966). Protein synthesis by cell–free extracts of Crithidia oncopelti. Bio–chemica et Biophysica Acta 114, 385–97.CrossRefGoogle ScholarPubMed
Clemens, M. J., Henshaw, E. C., Rahaminoff, H. & London, I. M. (1974). binding to 40S ribosomal subunits. A site for the regulation of initiation of protein synthesis by Hemin. Proceedings of the National Academy of Sciences USA 71, 2946–50.CrossRefGoogle ScholarPubMed
Cross, G. A. M. (1966). Protein synthesis in a cell–free system from Crithida oncopelti. Journal of General Microbiology 44, iii–iv.Google Scholar
Cross, G. A. M. (1975). Identification, purification and properties of clone specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Parasitology 71, 393–417.CrossRefGoogle ScholarPubMed
Hunt, T., Vanderhoff, G. & London, I.M. (1972). Control of globin synthesis: the role of Heme. Journal of Molecular Biology 66, 471–81.CrossRefGoogle ScholarPubMed
Kahan, D., Zahalsky, A. C. & Hunter, S. H. (1968). Protein synthesis in cell–free preparations of Crithidia fasciculata. Journal of Protozoology 15, 385–90.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lanham, S. M. & Godfrey, D. G. (1970). Isolation of salivarian trypanosomes from man and other mammals using DEAE–cellulose. Experimental Parasitology 28, 521–34.CrossRefGoogle ScholarPubMed
Lanham, S. M. & Taylor, A.E.R. (1972). Some properties of the immunogens (protective antigens) of a single variant of Trypanosoma brucei brucei. Journal of General Microbiology. 72, 101–16.CrossRefGoogle ScholarPubMed
Lee, S. Y., Mendecki, J. & Brawerman, G. (1971). A polynucleotide segment rich in adenylic acid in the rapidly–labelled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proceedings of the National Academy of Sciences USA 68, 1331–5.CrossRefGoogle Scholar
Pelham, H. R. B. & Jackson, R.J. (1976). An efficient mRNA–dependent transalation system from reticulocyte lysates. European Journal of Biochemistry 67, 247–56.CrossRefGoogle Scholar
Sheldon, R. M., Jurale, C. & Kates, J. (1972). Detection of polyadenylic acid sequences in viral and eukaryotic RNA. Proceedings of the National Academy of Sciences USA 69, 417–21.CrossRefGoogle ScholarPubMed
Sherman, I. R. (1976). The ribosomes of the simian malaria Plasmodium knowlesi. II. A cell–free protein synthesizing system. Comparative Biochemistry and Physiology 53B, 447–50.Google Scholar
Stuart, K. (1971). Evidence for the retention of kinetoplast DNA in an acriflavine–induced dyskinetoplastic strain of Trypanosoma brucei which replicates the altered central element of the kinetoplast. Journal of Cell Biology 49, 189–95.CrossRefGoogle Scholar
Vickerman, K. (1969). On the surface coat and flagellar adhesion in trypanosomes. Journal of Cell Science 5, 163–93.CrossRefGoogle ScholarPubMed
Williamson, J. & Brown, K.N. (1964). The chemical composition of trypanosomes. III. Antigenic constituents of Brucei trypanosomes. Experimental Parasitology 15, 46–68.CrossRefGoogle ScholarPubMed