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. 1988 Feb 25;16(4):1393–1406. doi: 10.1093/nar/16.4.1393

A head-to-tail tandem organization of hsp70 genes in Trypanosoma cruzi.

J M Requena 1, M C López 1, A Jimenez-Ruiz 1, J C de la Torre 1, C Alonso 1
PMCID: PMC336323  PMID: 2831499

Abstract

We describe the isolation and characterization of the T. cruzi hsp 70 DNA coding region which was found to be formed by multigene copies organized in a tandem array in a head-to-tail manner. The restriction pattern of one of the repetition units within the largest clone obtained from the genomic library, clone Tc70.6, shows that the hsp70 coding region should be formed by at least seven identical copies of 2.5 kb. We have found, however, the presence of restriction polymorphisms (Pvu II) within these repeats. Subsequent analysis of the time course of nuclear DNA digestion has revealed that the copy number per haploid genome could be as greater as 10. The analysis of the DNA and amino acid sequence of a fragment (70%) of one of the repetition units has shown the existence of a high homology with all the hsp70 genes of other organisms. The protein sequence homology of the fragment analyzed is as high as 88% when compared with that of the T. brucei hsp70. On the other hand, there are significant restriction site variations between both. The T. cruzi hsp70 contains at the C-terminal end a tetrapeptide repeat of the structure (GMPG)9.

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Selected References

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

  1. Ardeshir F., Flint J. E., Richman S. J., Reese R. T. A 75 kd merozoite surface protein of Plasmodium falciparum which is related to the 70 kd heat-shock proteins. EMBO J. 1987 Feb;6(2):493–499. doi: 10.1002/j.1460-2075.1987.tb04780.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bianco A. E., Favaloro J. M., Burkot T. R., Culvenor J. G., Crewther P. E., Brown G. V., Anders R. F., Coppel R. L., Kemp D. J. A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8713–8717. doi: 10.1073/pnas.83.22.8713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borst P. Discontinuous transcription and antigenic variation in trypanosomes. Annu Rev Biochem. 1986;55:701–732. doi: 10.1146/annurev.bi.55.070186.003413. [DOI] [PubMed] [Google Scholar]
  4. Chappell T. G., Welch W. J., Schlossman D. M., Palter K. B., Schlesinger M. J., Rothman J. E. Uncoating ATPase is a member of the 70 kilodalton family of stress proteins. Cell. 1986 Apr 11;45(1):3–13. doi: 10.1016/0092-8674(86)90532-5. [DOI] [PubMed] [Google Scholar]
  5. Dragon E. A., Sias S. R., Kato E. A., Gabe J. D. The genome of Trypanosoma cruzi contains a constitutively expressed, tandemly arranged multicopy gene homologous to a major heat shock protein. Mol Cell Biol. 1987 Mar;7(3):1271–1275. doi: 10.1128/mcb.7.3.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glass D. J., Polvere R. I., Van der Ploeg L. H. Conserved sequences and transcription of the hsp70 gene family in Trypanosoma brucei. Mol Cell Biol. 1986 Dec;6(12):4657–4666. doi: 10.1128/mcb.6.12.4657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Haltiner M., Kempe T., Tjian R. A novel strategy for constructing clustered point mutations. Nucleic Acids Res. 1985 Feb 11;13(3):1015–1025. doi: 10.1093/nar/13.3.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hedstrom R., Culpepper J., Harrison R. A., Agabian N., Newport G. A major immunogen in Schistosoma mansoni infections is homologous to the heat-shock protein Hsp70. J Exp Med. 1987 May 1;165(5):1430–1435. doi: 10.1084/jem.165.5.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hunt C., Morimoto R. I. Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6455–6459. doi: 10.1073/pnas.82.19.6455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hunter K. W., Cook C. L., Hayunga E. G. Leishmanial differentiation in vitro: induction of heat shock proteins. Biochem Biophys Res Commun. 1984 Dec 14;125(2):755–760. doi: 10.1016/0006-291x(84)90603-x. [DOI] [PubMed] [Google Scholar]
  13. Ingolia T. D., Craig E. A., McCarthy B. J. Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions. Cell. 1980 Oct;21(3):669–679. doi: 10.1016/0092-8674(80)90430-4. [DOI] [PubMed] [Google Scholar]
  14. Kochan J., Perkins M., Ravetch J. V. A tandemly repeated sequence determines the binding domain for an erythrocyte receptor binding protein of P. falciparum. Cell. 1986 Mar 14;44(5):689–696. doi: 10.1016/0092-8674(86)90834-2. [DOI] [PubMed] [Google Scholar]
  15. Korneluk R. G., Quan F., Gravel R. A. Rapid and reliable dideoxy sequencing of double-stranded DNA. Gene. 1985;40(2-3):317–323. doi: 10.1016/0378-1119(85)90055-1. [DOI] [PubMed] [Google Scholar]
  16. Lambowitz A. M., Kobayashi G. S., Painter A., Medoff G. Possible relationship of morphogenesis in pathogenic fungus, Histoplasma capsulatum, to heat shock response. Nature. 1983 Jun 30;303(5920):806–808. doi: 10.1038/303806a0. [DOI] [PubMed] [Google Scholar]
  17. Lancillotti F., Lopez M. C., Alonso C., Stollar B. D. Locations of Z-DNA in polytene chromosomes. J Cell Biol. 1985 May;100(5):1759–1766. doi: 10.1083/jcb.100.5.1759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lawrence F., Robert-Gero M. Induction of heat shock and stress proteins in promastigotes of three Leishmania species. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4414–4417. doi: 10.1073/pnas.82.13.4414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  20. Livak K. J., Freund R., Schweber M., Wensink P. C., Meselson M. Sequence organization and transcription at two heat shock loci in Drosophila. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5613–5617. doi: 10.1073/pnas.75.11.5613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Stahl H. D., Kemp D. J., Crewther P. E., Scanlon D. B., Woodrow G., Brown G. V., Bianco A. E., Anders R. F., Coppel R. L. Sequence of a cDNA encoding a small polymorphic histidine- and alanine-rich protein from Plasmodium falciparum. Nucleic Acids Res. 1985 Nov 11;13(21):7837–7846. doi: 10.1093/nar/13.21.7837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Van der Ploeg L. H. Discontinuous transcription and splicing in trypanosomes. Cell. 1986 Nov 21;47(4):479–480. doi: 10.1016/0092-8674(86)90608-2. [DOI] [PubMed] [Google Scholar]
  26. Van der Ploeg L. H., Giannini S. H., Cantor C. R. Heat shock genes: regulatory role for differentiation in parasitic protozoa. Science. 1985 Jun 21;228(4706):1443–1446. doi: 10.1126/science.4012301. [DOI] [PubMed] [Google Scholar]
  27. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]
  28. Zingales B., Colli W. Trypanosoma cruzi: interaction with host cells. Curr Top Microbiol Immunol. 1985;117:129–152. doi: 10.1007/978-3-642-70538-0_7. [DOI] [PubMed] [Google Scholar]

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