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. 1994 May 11;22(9):1663–1669. doi: 10.1093/nar/22.9.1663

Interaction of small ribosomal and transfer RNAs with a protein from Leishmania donovani.

A Ghosh 1, T Ghosh 1, S Ghosh 1, S Das 1, S Adhya 1
PMCID: PMC308046  PMID: 8202369

Abstract

Using synthetic antisense RNA from the 5'-untranslated region of the beta-tubulin gene as probe in gel retardation assays, a heat stable RNA-binding factor was identified in promastigotes of the kinetoplastid protozoan Leishmania donovani. The same or similar factors interact with several small ribosomal RNA (srRNA) species and, more weakly, with tRNA, as shown by binding and competition experiments. Deletion analysis indicated involvement of repeated purine-rich motifs on the antisense RNA, in the reaction. Related, conserved motifs occur on at least two of the srRNAs. By a modified Western blot assay, the RNA-binding species was identified as a single, small polypeptide. The activity is apparently specific for the promastigote stage of the parasite, being undetectable in amastigotes. The properties of this RNA-binding factor suggest that it is a novel, previously uncharacterized protein.

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Selected References

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  1. Bellofatto V., Cross G. A. Characterization of RNA transcripts from the alpha tubulin gene cluster of Leptomonas seymouri. Nucleic Acids Res. 1988 Apr 25;16(8):3455–3469. doi: 10.1093/nar/16.8.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bhaumik M., Das S., Adhya S. Evidence for translational control of beta-tubulin synthesis during differentiation of Leishmania donovani. Parasitology. 1991 Oct;103(Pt 2):197–205. doi: 10.1017/s0031182000059485. [DOI] [PubMed] [Google Scholar]
  3. Brun R., Krassner S. M. Quantitative ultrastructural investigations of mitochondrial development in Leishmania donovani during transformation. J Protozool. 1976 Nov;23(4):493–497. doi: 10.1111/j.1550-7408.1976.tb03824.x. [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. Cordingley J. S., Turner M. J. 6.5 S RNA; preliminary characterisation of unusual small RNAs in Trypanosoma brucei. Mol Biochem Parasitol. 1980 Apr;1(2):91–96. doi: 10.1016/0166-6851(80)90003-1. [DOI] [PubMed] [Google Scholar]
  6. Fong D., Chang K. P. Tubulin biosynthesis in the developmental cycle of a parasitic protozoan, Leishmania mexicana: changes during differentiation of motile and nonmotile stages. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7624–7628. doi: 10.1073/pnas.78.12.7624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fong D., Wallach M., Keithly J., Melera P. W., Chang K. P. Differential expression of mRNAs for alpha- and beta-tubulin during differentiation of the parasitic protozoan Leishmania mexicana. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5782–5786. doi: 10.1073/pnas.81.18.5782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gray M. W. Unusual pattern of ribonucleic acid components in the ribosome of Crithidia fasciculata, a trypanosomatid protozoan. Mol Cell Biol. 1981 Apr;1(4):347–357. doi: 10.1128/mcb.1.4.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harris M. E., Moore D. R., Hajduk S. L. Addition of uridines to edited RNAs in trypanosome mitochondria occurs independently of transcription. J Biol Chem. 1990 Jul 5;265(19):11368–11376. [PubMed] [Google Scholar]
  10. Hasan G., Turner M. J., Cordingley J. S. Ribosomal RNA genes of Trypanosoma brucei: mapping the regions specifying the six small ribosomal RNAs. Gene. 1984 Jan;27(1):75–86. doi: 10.1016/0378-1119(84)90240-3. [DOI] [PubMed] [Google Scholar]
  11. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  12. Hernández R., Nava G., Castañeda M. Small-size ribosomal RNA species in Trypanosoma cruzi. Mol Biochem Parasitol. 1983 Aug;8(4):297–304. doi: 10.1016/0166-6851(83)90076-2. [DOI] [PubMed] [Google Scholar]
  13. Janovy J., Jr Respiratory changes accompanying Leishmania to leptomonad transformation in Leishmania donovani. Exp Parasitol. 1967 Feb;20(1):51–55. doi: 10.1016/0014-4894(67)90021-5. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Laird P. W. Trans splicing in trypanosomes--archaism or adaptation? Trends Genet. 1989 Jul;5(7):204–208. doi: 10.1016/0168-9525(89)90082-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Perry K. L., Watkins K. P., Agabian N. Trypanosome mRNAs have unusual "cap 4" structures acquired by addition of a spliced leader. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8190–8194. doi: 10.1073/pnas.84.23.8190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. RUDZINSKA M. A., D ALESANDRO P. A., TRAGER W. THE FINE STRUCTURE OF LEISHMANIA DONOVANI AND THE ROLE OF THE KINETOPLAST IN THE LEISHMANI-LEPTOMONAD TRANSFORMATION. J Protozool. 1964 May;11:166–191. doi: 10.1111/j.1550-7408.1964.tb01739.x. [DOI] [PubMed] [Google Scholar]
  18. Schnare M. N., Spencer D. F., Gray M. W. Primary structures of four novel small ribosomal RNAs from Crithidia fasciculata. Can J Biochem Cell Biol. 1983 Jan;61(1):38–45. doi: 10.1139/o83-006. [DOI] [PubMed] [Google Scholar]
  19. Simpson A. M., Suyama Y., Dewes H., Campbell D. A., Simpson L. Kinetoplastid mitochondria contain functional tRNAs which are encoded in nuclear DNA and also contain small minicircle and maxicircle transcripts of unknown function. Nucleic Acids Res. 1989 Jul 25;17(14):5427–5445. doi: 10.1093/nar/17.14.5427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Simpson L. The leishmania-leptomonad transformation of Leishmania donovani: nutritional requirements, respiration changes and antigenic changes. J Protozool. 1968 Feb;15(1):201–207. doi: 10.1111/j.1550-7408.1968.tb02112.x. [DOI] [PubMed] [Google Scholar]
  21. Specht T., Wolters J., Erdmann V. A. Compilation of 5S rRNA and 5S rRNA gene sequences. Nucleic Acids Res. 1990 Apr 25;18 (Suppl):2215–2230. doi: 10.1093/nar/18.suppl.2215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sprinzl M., Hartmann T., Meissner F., Moll J., Vorderwülbecke T. Compilation of tRNA sequences and sequences of tRNA genes. Nucleic Acids Res. 1987;15 (Suppl):r53–188. doi: 10.1093/nar/15.suppl.r53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Veldman G. M., Klootwijk J., de Regt V. C., Planta R. J., Branlant C., Krol A., Ebel J. P. The primary and secondary structure of yeast 26S rRNA. Nucleic Acids Res. 1981 Dec 21;9(24):6935–6952. doi: 10.1093/nar/9.24.6935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wallach M., Fong D., Chang K. P. Post-transcriptional control of tubulin biosynthesis during leishmanial differentiation. Nature. 1982 Oct 14;299(5884):650–652. doi: 10.1038/299650a0. [DOI] [PubMed] [Google Scholar]
  25. White T. C., Rudenko G., Borst P. Three small RNAs within the 10 kb trypanosome rRNA transcription unit are analogous to domain VII of other eukaryotic 28S rRNAs. Nucleic Acids Res. 1986 Dec 9;14(23):9471–9489. doi: 10.1093/nar/14.23.9471. [DOI] [PMC free article] [PubMed] [Google Scholar]

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