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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Aug;80(15):4634–4638. doi: 10.1073/pnas.80.15.4634

Tubulin genes of Trypanosoma brucei: a tightly clustered family of alternating genes.

T Seebeck, P A Whittaker, M A Imboden, N Hardman, R Braun
PMCID: PMC384098  PMID: 6308633

Abstract

African trypanosomes are the causative agents of many medically and economically important diseases of man and domestic animals. The cell body of these blood-dwelling protozoa is enveloped with a dense layer of pellicular microtubules, which confer both motility and mechanical stability on these cells; microtubules are also important components of the flagellum. The major structural components of the microtubuli are two related proteins, alpha- and beta-tubulin. We have analyzed the genomic organization of alpha- and beta-tubulin genes in Trypanosoma brucei. In this organism, the majority of these genes are arranged in a single, tightly packed cluster of alternating alpha- and beta-tubulin genes with a basic repeat length together of 3.6 kilobase pairs. A genomic library of T. brucei was constructed by using the phage vector A 1059, and recombinant phages carrying tubulin genes were isolated by screening the library with heterologous chicken tubulin cDNA probes. The results of restriction endonuclease and hybridization analysis of DNA isolated from recombinant phages, and subcloned fragments thereof, were compatible with the restriction maps derived from digestion and Southern blot hybridization of genomic DNA.

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Selected References

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

  1. Agabian N., Thomashow L., Milhausen M., Stuart K. Structural analysis of variant and invariant genes in trypanosomes. Am J Trop Med Hyg. 1980 Sep;29(5 Suppl):1043–1049. doi: 10.4269/ajtmh.1980.29.1043. [DOI] [PubMed] [Google Scholar]
  2. Alexandraki D., Ruderman J. V. Sequence heterogeneity, multiplicity, and genomic organization of alpha- and beta-tubulin genes in sea urchins. Mol Cell Biol. 1981 Dec;1(12):1125–1137. doi: 10.1128/mcb.1.12.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borst P., van der Ploeg M., van Hoek J. F., Tas J., James J. On the DNA content and ploidy of trypanosomes. Mol Biochem Parasitol. 1982 Jul;6(1):13–23. doi: 10.1016/0166-6851(82)90049-4. [DOI] [PubMed] [Google Scholar]
  4. Brun R., Schönenberger Cultivation and in vitro cloning or procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Short communication. Acta Trop. 1979 Sep;36(3):289–292. [PubMed] [Google Scholar]
  5. Cleveland D. W., Hughes S. H., Stubblefield E., Kirschner M. W., Varmus H. E. Multiple alpha and beta tubulin genes represent unlinked and dispersed gene families. J Biol Chem. 1981 Mar 25;256(6):3130–3134. [PubMed] [Google Scholar]
  6. Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
  7. Jaggi R. B., Wyler T., Ryffel G. U. Comparative analysis of Xenopus tropicalis and Xenopus laevis vitellogenin gene sequences. Nucleic Acids Res. 1982 Mar 11;10(5):1515–1533. doi: 10.1093/nar/10.5.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kalfayan L., Wensink P. C. alpha-Tubulin genes of Drosophila. Cell. 1981 Apr;24(1):97–106. doi: 10.1016/0092-8674(81)90505-5. [DOI] [PubMed] [Google Scholar]
  9. Karn J., Brenner S., Barnett L., Cesareni G. Novel bacteriophage lambda cloning vector. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5172–5176. doi: 10.1073/pnas.77.9.5172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laird C. D., McConaughy B. L., McCarthy B. J. Rate of fixation of nucleotide substitutions in evolution. Nature. 1969 Oct 11;224(5215):149–154. doi: 10.1038/224149a0. [DOI] [PubMed] [Google Scholar]
  11. Lemischka I. R., Farmer S., Racaniello V. R., Sharp P. A. Nucleotide sequence and evolution of a mammalian alpha-tubulin messenger RNA. J Mol Biol. 1981 Sep 5;151(1):101–120. doi: 10.1016/0022-2836(81)90223-0. [DOI] [PubMed] [Google Scholar]
  12. Lemischka I., Sharp P. A. The sequences of an expressed rat alpha-tubulin gene and a pseudogene with an inserted repetitive element. Nature. 1982 Nov 25;300(5890):330–335. doi: 10.1038/300330a0. [DOI] [PubMed] [Google Scholar]
  13. Luduena R. F., Woodward D. O. Isolation and partial characterization of alpha and beta-tubulin from outer doublets of sea-urchin sperm and microtubules of chick-embryo brain. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3594–3598. doi: 10.1073/pnas.70.12.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ormerod W. E. Human and animal trypanosomiases as world public health problems. Pharmacol Ther. 1979;6(1):1–40. doi: 10.1016/0163-7258(79)90055-x. [DOI] [PubMed] [Google Scholar]
  15. Ponstingl H., Krauhs E., Little M., Kempf T. Complete amino acid sequence of alpha-tubulin from porcine brain. Proc Natl Acad Sci U S A. 1981 May;78(5):2757–2761. doi: 10.1073/pnas.78.5.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
  17. Snyder J. A., McIntosh J. R. Biochemistry and physiology of microtubules. Annu Rev Biochem. 1976;45:699–720. doi: 10.1146/annurev.bi.45.070176.003411. [DOI] [PubMed] [Google Scholar]
  18. Sánchez F., Natzle J. E., Cleveland D. W., Kirschner M. W., McCarthy B. J. A dispersed multigene family encoding tubulin in Drosophila melanogaster. Cell. 1980 Dec;22(3):845–854. doi: 10.1016/0092-8674(80)90561-9. [DOI] [PubMed] [Google Scholar]
  19. Thomashow L. S., Milhausen M., Rutter W. J., Agabian N. Tubulin genes are tandemly linked and clustered in the genome of trypanosoma brucei. Cell. 1983 Jan;32(1):35–43. doi: 10.1016/0092-8674(83)90494-4. [DOI] [PubMed] [Google Scholar]
  20. Ullman J. S., McCarthy B. J. The relationship between mismatched base pairs and the thermal stability of DNA duplexes. I. Effects of depurination and chain scission. Biochim Biophys Acta. 1973 Feb 4;294(1):405–415. doi: 10.1016/0005-2787(73)90095-6. [DOI] [PubMed] [Google Scholar]
  21. Valenzuela P., Quiroga M., Zaldivar J., Rutter W. J., Kirschner M. W., Cleveland D. W. Nucleotide and corresponding amino acid sequences encoded by alpha and beta tubulin mRNAs. Nature. 1981 Feb 19;289(5799):650–655. doi: 10.1038/289650a0. [DOI] [PubMed] [Google Scholar]
  22. Wilde C. D., Crowther C. E., Cowan N. J. Diverse mechanisms in the generation of human beta-tubulin pseudogenes. Science. 1982 Aug 6;217(4559):549–549. doi: 10.1126/science.6178164. [DOI] [PubMed] [Google Scholar]
  23. Wilde C. D., Crowther C. E., Cowan N. J. Isolation of a multigene family containing human alpha-tubulin sequences. J Mol Biol. 1982 Mar 15;155(4):533–538. doi: 10.1016/0022-2836(82)90486-7. [DOI] [PubMed] [Google Scholar]
  24. Wilde C. D., Crowther C. E., Cripe T. P., Gwo-Shu Lee M., Cowan N. J. Evidence that a human beta-tubulin pseudogene is derived from its corresponding mRNA. Nature. 1982 May 6;297(5861):83–84. doi: 10.1038/297083a0. [DOI] [PubMed] [Google Scholar]

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