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. 1992 May;12(5):2043–2049. doi: 10.1128/mcb.12.5.2043

Guide RNAs for transcripts with developmentally regulated RNA editing are present in both life cycle stages of Trypanosoma brucei.

D J Koslowsky 1, G R Riley 1, J E Feagin 1, K Stuart 1
PMCID: PMC364375  PMID: 1373804

Abstract

RNA editing of several mitochondrial transcripts in Trypanosoma brucei is developmentally regulated. The cytochrome b and cytochrome oxidase II mRNAs are edited in procyclic-form parasites but are primarily unedited in bloodstream forms. The latter forms lack the mitochondrial respiratory system present in procyclic forms. Editing of the NADH dehydrogenase 7 (ND7) and ND8 transcripts is also developmentally regulated but occurs preferentially in bloodstream forms. Other transcripts, cytochrome oxidase III and ATPase 6, are edited in both life forms. We have identified many minicircle-encoded guide RNAs (gRNAs) for ATPase 6, ND7, and ND8. The characteristics of these gRNAs reveal how extensively edited RNA can be edited in the 3'-to-5' direction. Northern (RNA) blot and primer extension analyses indicate that gRNAs for transcripts whose editing is developmentally regulated are present in both procyclic and bloodstream form parasites. These results suggest that the developmental regulation of editing in these transcripts is not controlled by the presence or absence of gRNAs.

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Selected References

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  1. Barrois M., Riou G., Galibert F. Complete nucleotide sequence of minicircle kinetoplast DNA from Trypanosoma equiperdum. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3323–3327. doi: 10.1073/pnas.78.6.3323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bhat G. J., Koslowsky D. J., Feagin J. E., Smiley B. L., Stuart K. An extensively edited mitochondrial transcript in kinetoplastids encodes a protein homologous to ATPase subunit 6. Cell. 1990 Jun 1;61(5):885–894. doi: 10.1016/0092-8674(90)90199-o. [DOI] [PubMed] [Google Scholar]
  3. Blum B., Bakalara N., Simpson L. A model for RNA editing in kinetoplastid mitochondria: "guide" RNA molecules transcribed from maxicircle DNA provide the edited information. Cell. 1990 Jan 26;60(2):189–198. doi: 10.1016/0092-8674(90)90735-w. [DOI] [PubMed] [Google Scholar]
  4. Blum B., Simpson L. Guide RNAs in kinetoplastid mitochondria have a nonencoded 3' oligo(U) tail involved in recognition of the preedited region. Cell. 1990 Jul 27;62(2):391–397. doi: 10.1016/0092-8674(90)90375-o. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Englund P. T., Hajduk S. L., Marini J. C. The molecular biology of trypanosomes. Annu Rev Biochem. 1982;51:695–726. doi: 10.1146/annurev.bi.51.070182.003403. [DOI] [PubMed] [Google Scholar]
  7. Feagin J. E., Abraham J. M., Stuart K. Extensive editing of the cytochrome c oxidase III transcript in Trypanosoma brucei. Cell. 1988 May 6;53(3):413–422. doi: 10.1016/0092-8674(88)90161-4. [DOI] [PubMed] [Google Scholar]
  8. Feagin J. E., Jasmer D. P., Stuart K. Apocytochrome b and other mitochondrial DNA sequences are differentially expressed during the life cycle of Trypanosoma brucei. Nucleic Acids Res. 1985 Jun 25;13(12):4577–4596. doi: 10.1093/nar/13.12.4577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feagin J. E., Jasmer D. P., Stuart K. Developmentally regulated addition of nucleotides within apocytochrome b transcripts in Trypanosoma brucei. Cell. 1987 May 8;49(3):337–345. doi: 10.1016/0092-8674(87)90286-8. [DOI] [PubMed] [Google Scholar]
  10. Feagin J. E., Jasmer D. P., Stuart K. Differential mitochondrial gene expression between slender and stumpy bloodforms of Trypanosoma brucei. Mol Biochem Parasitol. 1986 Sep;20(3):207–214. doi: 10.1016/0166-6851(86)90100-3. [DOI] [PubMed] [Google Scholar]
  11. Feagin J. E., Stuart K. Developmental aspects of uridine addition within mitochondrial transcripts of Trypanosoma brucei. Mol Cell Biol. 1988 Mar;8(3):1259–1265. doi: 10.1128/mcb.8.3.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feagin J. E., Stuart K. Differential expression of mitochondrial genes between life cycle stages of Trypanosoma brucei. Proc Natl Acad Sci U S A. 1985 May;82(10):3380–3384. doi: 10.1073/pnas.82.10.3380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Jasmer D. P., Feagin J. E., Stuart K. Diverse patterns of expression of the cytochrome c oxidase subunit I gene and unassigned reading frames 4 and 5 during the life cycle of Trypanosoma brucei. Mol Cell Biol. 1985 Nov;5(11):3041–3047. doi: 10.1128/mcb.5.11.3041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jasmer D. P., Stuart K. Conservation of kinetoplastid minicircle characteristics without nucleotide sequence conservation. Mol Biochem Parasitol. 1986 Mar;18(3):257–269. doi: 10.1016/0166-6851(86)90084-8. [DOI] [PubMed] [Google Scholar]
  16. Jasmer D. P., Stuart K. Sequence organization in African trypanosome minicircles is defined by 18 base pair inverted repeats. Mol Biochem Parasitol. 1986 Mar;18(3):321–331. doi: 10.1016/0166-6851(86)90089-7. [DOI] [PubMed] [Google Scholar]
  17. Koslowsky D. J., Bhat G. J., Perrollaz A. L., Feagin J. E., Stuart K. The MURF3 gene of T. brucei contains multiple domains of extensive editing and is homologous to a subunit of NADH dehydrogenase. Cell. 1990 Sep 7;62(5):901–911. doi: 10.1016/0092-8674(90)90265-g. [DOI] [PubMed] [Google Scholar]
  18. Koslowsky D. J., Bhat G. J., Read L. K., Stuart K. Cycles of progressive realignment of gRNA with mRNA in RNA editing. Cell. 1991 Nov 1;67(3):537–546. doi: 10.1016/0092-8674(91)90528-7. [DOI] [PubMed] [Google Scholar]
  19. Michelotti E. F., Hajduk S. L. Developmental regulation of trypanosome mitochondrial gene expression. J Biol Chem. 1987 Jan 15;262(2):927–932. [PubMed] [Google Scholar]
  20. Pollard V. W., Hajduk S. L. Trypanosoma equiperdum minicircles encode three distinct primary transcripts which exhibit guide RNA characteristics. Mol Cell Biol. 1991 Mar;11(3):1668–1675. doi: 10.1128/mcb.11.3.1668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pollard V. W., Rohrer S. P., Michelotti E. F., Hancock K., Hajduk S. L. Organization of minicircle genes for guide RNAs in Trypanosoma brucei. Cell. 1990 Nov 16;63(4):783–790. doi: 10.1016/0092-8674(90)90144-4. [DOI] [PubMed] [Google Scholar]
  22. Souza A. E., Myler P. J., Stuart K. Maxicircle CR1 transcripts of Trypanosoma brucei are edited and developmentally regulated and encode a putative iron-sulfur protein homologous to an NADH dehydrogenase subunit. Mol Cell Biol. 1992 May;12(5):2100–2107. doi: 10.1128/mcb.12.5.2100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stuart K. D., Gelvin S. B. Localization of kinetoplast DNA maxicircle transcripts in bloodstream and procyclic form Trypanosoma brucei. Mol Cell Biol. 1982 Jul;2(7):845–852. doi: 10.1128/mcb.2.7.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stuart K., Gobright E., Jenni L., Milhausen M., Thomashow L., Agabian N. The IsTaR 1 serodeme of Trypanosoma brucei: development of a new serodeme. J Parasitol. 1984 Oct;70(5):747–754. [PubMed] [Google Scholar]
  25. Stuart K. Kinetoplast DNA, mitochondrial DNA with a difference. Mol Biochem Parasitol. 1983 Oct;9(2):93–104. doi: 10.1016/0166-6851(83)90103-2. [DOI] [PubMed] [Google Scholar]
  26. Stuart K. Mitochondrial DNA of an African trypanosome. J Cell Biochem. 1983;23(1-4):13–26. doi: 10.1002/jcb.240230103. [DOI] [PubMed] [Google Scholar]
  27. Stuart K. RNA editing in mitochondrial mRNA of trypanosomatids. Trends Biochem Sci. 1991 Feb;16(2):68–72. doi: 10.1016/0968-0004(91)90027-s. [DOI] [PubMed] [Google Scholar]
  28. Stuart K. RNA editing in trypanosomatid mitochondria. Annu Rev Microbiol. 1991;45:327–344. doi: 10.1146/annurev.mi.45.100191.001551. [DOI] [PubMed] [Google Scholar]
  29. Sturm N. R., Simpson L. Kinetoplast DNA minicircles encode guide RNAs for editing of cytochrome oxidase subunit III mRNA. Cell. 1990 Jun 1;61(5):879–884. doi: 10.1016/0092-8674(90)90198-n. [DOI] [PubMed] [Google Scholar]
  30. Vickerman K. Developmental cycles and biology of pathogenic trypanosomes. Br Med Bull. 1985 Apr;41(2):105–114. doi: 10.1093/oxfordjournals.bmb.a072036. [DOI] [PubMed] [Google Scholar]
  31. van der Spek H., Arts G. J., Zwaal R. R., van den Burg J., Sloof P., Benne R. Conserved genes encode guide RNAs in mitochondria of Crithidia fasciculata. EMBO J. 1991 May;10(5):1217–1224. doi: 10.1002/j.1460-2075.1991.tb08063.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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