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. 1978 Sep;5(9):3171–3182. doi: 10.1093/nar/5.9.3171

A characterization of mRNA activites and their sequence complexities in Trypanosoma brucei: partial purification and properties of the VSSA mRNA.

R O Williams, K B Marcu, J R Young, L Rovis, S C Williams
PMCID: PMC342239  PMID: 704349

Abstract

Polyadenylated RNA isolated from a clone of Trypanosoma brucei was shown to direct the synthesis of a variety of polypeptides in a cell-free system. A predominant 58,000 dalton polypeptide was immunoprecipitated with antisera to the T. brucei variant specific surface antigen (VSSA). The mRNA that directed the synthesis of the VSSA was 2.0 kilobases (kb) long as measured by polyacrylamide gel electrophoresis in 98% formamide. Complementary DNA was prepared with avian myeloblastosis virus reverse transcriptase and the nucleotide sequence complexities of the total polysomal poly(A)+RNA and a gel purified VSSA mRNA were measured. 20% of the total cellular poly(A)+RNA contained abundant sequences with an apparent complexity of 9.6 kb; 42% of the purified VSSA mRNA contained abundant sequences with a complexity of 7.2 kb. Complementary DNA synthesized from gel purified VSSA mRNA was hybridized to total cellular poly(A)+RNA isolated from an unrelated T. brucei clone expressing a different variant antigen. A portion of the low complexity RNA sequence component was absent in the heterologous mRNA population but the same plateau of hybridization was achieved (93%). The abundance of some of the low complexity mRNAs appears to be T. brucei clone specific.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baltz T., Baltz D., Pautrizel R., Richet C., Lamblin G., Degand P. Chemical and immunological characterization of specific glycoproteins from Trypanosoma equiperdum variants. FEBS Lett. 1977 Oct 1;82(1):93–96. doi: 10.1016/0014-5793(77)80893-4. [DOI] [PubMed] [Google Scholar]
  2. Bridgen P. J., Cross G. A., Bridgen J. N-terminal amino acid sequences of variant-specific surface antigens from Trypanosoma brucei. Nature. 1976 Oct 14;263(5578):613–614. doi: 10.1038/263613a0. [DOI] [PubMed] [Google Scholar]
  3. Brownlee G. G., Cartwright E. M., Cowan N. J., Jarvis J. M., Milstein C. Purification and sequence of messenger RNA for immunoglobulin light chains. Nat New Biol. 1973 Aug 22;244(138):236–240. doi: 10.1038/newbio244236a0. [DOI] [PubMed] [Google Scholar]
  4. Cowan N. J., Secher D. S., Milstein C. Purification and sequence analysis of the mRNA coding for an immunoglobulin heavy chain. Eur J Biochem. 1976 Jan 15;61(2):353–368. doi: 10.1111/j.1432-1033.1976.tb10029.x. [DOI] [PubMed] [Google Scholar]
  5. Cross G. A. Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Parasitology. 1975 Dec;71(3):393–417. doi: 10.1017/s003118200004717x. [DOI] [PubMed] [Google Scholar]
  6. Doyle J. J., Behin R., Mauel J., Rowe D. S. Antibody-induced movement of protozoan surface membrane antigens. Ann N Y Acad Sci. 1975 Jun 30;254:315–325. doi: 10.1111/j.1749-6632.1975.tb29181.x. [DOI] [PubMed] [Google Scholar]
  7. Duesberg P. H., Vogt P. K. Gel electrophoresis of avian leukosis and sarcoma viral RNA in formamide: comparison with other viral and cellular RNA species. J Virol. 1973 Sep;12(3):594–599. doi: 10.1128/jvi.12.3.594-599.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Efstratiadis A., Kafatos F. C., Maxam A. M., Maniatis T. Enzymatic in vitro synthesis of globin genes. Cell. 1976 Feb;7(2):279–288. doi: 10.1016/0092-8674(76)90027-1. [DOI] [PubMed] [Google Scholar]
  9. Eggitt M. J., Tappenden L., Brown K. N. Translation in a reticulocyte cell-free system of RNA isolated from blood and culture forms of Trypanosoma brucei. Parasitology. 1977 Oct;75(2):133–141. doi: 10.1017/s0031182000062272. [DOI] [PubMed] [Google Scholar]
  10. Hastie N. D., Bishop J. O. The expression of three abundance classes of messenger RNA in mouse tissues. Cell. 1976 Dec;9(4 Pt 2):761–774. doi: 10.1016/0092-8674(76)90139-2. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lanham S. M., Godfrey D. G. Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose. Exp Parasitol. 1970 Dec;28(3):521–534. doi: 10.1016/0014-4894(70)90120-7. [DOI] [PubMed] [Google Scholar]
  13. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marcu K., Dudock B. Characterization of a highly efficient protein synthesizing system derived from commercial wheat germ. Nucleic Acids Res. 1974 Nov;1(11):1385–1397. doi: 10.1093/nar/1.11.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rovis L., Barbet A. F., Williams R. O. Characterisation of the surface coat of Trypanosoma congolense. Nature. 1978 Feb 16;271(5646):654–656. doi: 10.1038/271654a0. [DOI] [PubMed] [Google Scholar]
  17. Taylor D. W., Cross G. A. The synthesis of a variant-specific antigen by Trypanosoma brucei in vitro. Parasitology. 1977 Feb;74(1):47–60. doi: 10.1017/s0031182000047521. [DOI] [PubMed] [Google Scholar]
  18. Weeks D. P., Collis P. S. Induction of microtubule protein synthesis in Chlamydomonas reinhardi during flagellar regeneration. Cell. 1976 Sep;9(1):15–27. doi: 10.1016/0092-8674(76)90048-9. [DOI] [PubMed] [Google Scholar]

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