Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1987 Mar;7(3):1271–1275. doi: 10.1128/mcb.7.3.1271

The genome of Trypanosoma cruzi contains a constitutively expressed, tandemly arranged multicopy gene homologous to a major heat shock protein.

E A Dragon, S R Sias, E A Kato, J D Gabe
PMCID: PMC365202  PMID: 3550435

Abstract

cDNA libraries have been constructed in the plasmid vector pUC18 with mRNA isolated from both epimastigotes and trypomastigotes of the Peru strain of Trypanosoma cruzi. Pools of randomly selected clones were analyzed by hybridization-selection-translation. Translation products were immunoprecipitated either with normal human sera or with sera from patients with Chagas' disease (chagasic sera), and the immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With this approach, a cDNA clone (pEC5) was identified which encodes a portion of an 85,000-Mr polypeptide. A genomic clone was subsequently isolated (FG1) by using oligonucleotide probes derived from the DNA sequence of this cDNA clone. A portion of this clone was isolated and sequenced, and the coding region for the protein was identified. Computer analysis of the predicted protein sequence indicates that this protein is closely related to the 83,000-Mr heat shock protein (hsp83) of Drosophila melanogaster, the hsp90 of Saccharomyces cerevisiae, and the hsp90 of chicken. This gene is tandemly organized in the T. cruzi genome as a cluster of 6 to 10 copies.

Full text

PDF
1274

Images in this article

Selected References

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

  1. Andrews N. W., Katzin A. M., Colli W. Mapping of surface glycoproteins of Trypanosoma cruzi by two-dimensional electrophoresis. A correlation with the cell invasion capacity. Eur J Biochem. 1984 May 2;140(3):599–604. doi: 10.1111/j.1432-1033.1984.tb08144.x. [DOI] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beard C. A., Wrightsman R. A., Manning J. E. Identification of monoclonal antibodies against the trypomastigote stage of Trypanosoma cruzi by use of iminobiotinylated surface polypeptides. Mol Biochem Parasitol. 1985 Aug;16(2):199–212. doi: 10.1016/0166-6851(85)90087-8. [DOI] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Catelli M. G., Binart N., Feramisco J. R., Helfman D. M. Cloning of the chick hsp 90 cDNA in expression vector. Nucleic Acids Res. 1985 Sep 11;13(17):6035–6047. doi: 10.1093/nar/13.17.6035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dragon E. A., Brothers V. M., Wrightsman R. A., Manning J. A Mr 90 000 surface polypeptide of Trypanosoma cruzi as a candidate for a Chagas' disease diagnostic antigen. Mol Biochem Parasitol. 1985 Sep;16(3):213–229. doi: 10.1016/0166-6851(85)90065-9. [DOI] [PubMed] [Google Scholar]
  7. Farrelly F. W., Finkelstein D. B. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae. J Biol Chem. 1984 May 10;259(9):5745–5751. [PubMed] [Google Scholar]
  8. Gonzalez A., Lerner T. J., Huecas M., Sosa-Pineda B., Nogueira N., Lizardi P. M. Apparent generation of a segmented mRNA from two separate tandem gene families in Trypanosoma cruzi. Nucleic Acids Res. 1985 Aug 26;13(16):5789–5804. doi: 10.1093/nar/13.16.5789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  10. Hackett R. W., Lis J. T. Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus of D. melanogaster. Nucleic Acids Res. 1983 Oct 25;11(20):7011–7030. doi: 10.1093/nar/11.20.7011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  12. Hudson L., Hindmarsh P. J. The relationship between autoimmunity and Chagas' disease: causal or coincidental? Curr Top Microbiol Immunol. 1985;117:167–177. doi: 10.1007/978-3-642-70538-0_9. [DOI] [PubMed] [Google Scholar]
  13. Lanar D. E., Levy L. S., Manning J. E. Complexity and content of the DNA and RNA in Trypanosoma cruzi. Mol Biochem Parasitol. 1981 Sep;3(5):327–341. doi: 10.1016/0166-6851(81)90006-2. [DOI] [PubMed] [Google Scholar]
  14. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  15. Nogueira N., Chaplan S., Tydings J. D., Unkeless J., Cohn Z. Trypanosoma cruzi. Surface antigens of blood and culture forms. J Exp Med. 1981 Mar 1;153(3):629–639. doi: 10.1084/jem.153.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nogueira N., Unkeless J., Cohn Z. Specific glycoprotein antigens on the surface of insect and mammalian stages of Trypanosoma cruzi. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1259–1263. doi: 10.1073/pnas.79.4.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  18. Ricciardi R. P., Miller J. S., Roberts B. E. Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4927–4931. doi: 10.1073/pnas.76.10.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Snary D., Hudson L. Trypanosoma cruzi cell surface proteins: identification of one major glycoprotein. FEBS Lett. 1979 Apr 1;100(1):166–170. doi: 10.1016/0014-5793(79)81156-4. [DOI] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ullrich A., Shine J., Chirgwin J., Pictet R., Tischer E., Rutter W. J., Goodman H. M. Rat insulin genes: construction of plasmids containing the coding sequences. Science. 1977 Jun 17;196(4296):1313–1319. doi: 10.1126/science.325648. [DOI] [PubMed] [Google Scholar]
  23. Ullrich S. J., Robinson E. A., Law L. W., Willingham M., Appella E. A mouse tumor-specific transplantation antigen is a heat shock-related protein. Proc Natl Acad Sci U S A. 1986 May;83(10):3121–3125. doi: 10.1073/pnas.83.10.3121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES