Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1991 May;10(5):1217–1224. doi: 10.1002/j.1460-2075.1991.tb08063.x

Conserved genes encode guide RNAs in mitochondria of Crithidia fasciculata.

H van der Spek 1, G J Arts 1, R R Zwaal 1, J van den Burg 1, P Sloof 1, R Benne 1
PMCID: PMC452776  PMID: 1708723

Abstract

RNA editing is the post-transcriptional alteration of the nucleotide sequence of RNA, which in trypanosome mitochondria is characterized by the insertion and deletion of uridine residues. It has recently been proposed that the information for the sequence alteration in Leishmania tarentolae is provided by small guide (g) RNAs encoded in the mitochondrial DNA [Blum et al. (1990) Cell, 60, 189-198]. We are studying the mechanism of RNA editing in the insect trypanosome Crithidia fasciculata and report that: (i) a full length, conventional DNA gene or an independently replicating RNA gene that could encode the edited MURF3 transcript is absent when probed for in sensitive, calibrated assay systems; (ii) in all cases (seven) investigated in C. fasciculata so far, putative gRNA genes are found in a position in the mitochondrial DNA virtually identical to that in L. tarentolae and (iii) also in C. fasciculata, the putative gRNA genes are transcribed into small RNAs with discrete 5' ends. These results provide strong evolutionary evidence in support of the participation of gRNAs in RNA editing. Remarkably, in C. fasciculata the basepaired region of some putative gRNA:mRNA hybrids contains a C:A non-Watson-Crick basepair.

Full text

PDF
1217

Images in this article

Selected References

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

  1. Abraham J. M., Feagin J. E., Stuart K. Characterization of cytochrome c oxidase III transcripts that are edited only in the 3' region. Cell. 1988 Oct 21;55(2):267–272. doi: 10.1016/0092-8674(88)90049-9. [DOI] [PubMed] [Google Scholar]
  2. Benne R. RNA editing in trypanosomes: is there a message? Trends Genet. 1990 Jun;6(6):177–181. doi: 10.1016/0168-9525(90)90173-4. [DOI] [PubMed] [Google Scholar]
  3. Benne R. RNA-editing in trypanosome mitochondria. Biochim Biophys Acta. 1989 Mar 1;1007(2):131–139. doi: 10.1016/0167-4781(89)90031-6. [DOI] [PubMed] [Google Scholar]
  4. Benne R., Van den Burg J., Brakenhoff J. P., Sloof P., Van Boom J. H., Tromp M. C. Major transcript of the frameshifted coxII gene from trypanosome mitochondria contains four nucleotides that are not encoded in the DNA. Cell. 1986 Sep 12;46(6):819–826. doi: 10.1016/0092-8674(86)90063-2. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Birkenmeyer L., Ray D. S. Replication of kinetoplast DNA in isolated kinetoplasts from Crithidia fasciculata. Identification of minicircle DNA replication intermediates. J Biol Chem. 1986 Feb 15;261(5):2362–2368. [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Bogenhagen D. F., Brown D. D. Nucleotide sequences in Xenopus 5S DNA required for transcription termination. Cell. 1981 Apr;24(1):261–270. doi: 10.1016/0092-8674(81)90522-5. [DOI] [PubMed] [Google Scholar]
  10. Borst P., Hoeijmakers J. H. Kinetoplast DNA. Plasmid. 1979 Jan;2(1):20–40. doi: 10.1016/0147-619x(79)90003-9. [DOI] [PubMed] [Google Scholar]
  11. Cattaneo R., Kaelin K., Baczko K., Billeter M. A. Measles virus editing provides an additional cysteine-rich protein. Cell. 1989 Mar 10;56(5):759–764. doi: 10.1016/0092-8674(89)90679-x. [DOI] [PubMed] [Google Scholar]
  12. Chen S. H., Habib G., Yang C. Y., Gu Z. W., Lee B. R., Weng S. A., Silberman S. R., Cai S. J., Deslypere J. P., Rosseneu M. Apolipoprotein B-48 is the product of a messenger RNA with an organ-specific in-frame stop codon. Science. 1987 Oct 16;238(4825):363–366. doi: 10.1126/science.3659919. [DOI] [PubMed] [Google Scholar]
  13. Covello P. S., Gray M. W. RNA editing in plant mitochondria. Nature. 1989 Oct 19;341(6243):662–666. doi: 10.1038/341662a0. [DOI] [PubMed] [Google Scholar]
  14. Decker C. J., Sollner-Webb B. RNA editing involves indiscriminate U changes throughout precisely defined editing domains. Cell. 1990 Jun 15;61(6):1001–1011. doi: 10.1016/0092-8674(90)90065-m. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Gualberto J. M., Lamattina L., Bonnard G., Weil J. H., Grienenberger J. M. RNA editing in wheat mitochondria results in the conservation of protein sequences. Nature. 1989 Oct 19;341(6243):660–662. doi: 10.1038/341660a0. [DOI] [PubMed] [Google Scholar]
  18. Gualberto J. M., Weil J. H., Grienenberger J. M. Editing of the wheat coxIII transcript: evidence for twelve C to U and one U to C conversions and for sequence similarities around editing sites. Nucleic Acids Res. 1990 Jul 11;18(13):3771–3776. doi: 10.1093/nar/18.13.3771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Hensgens L. A., Brakenhoff J., De Vries B. F., Sloof P., Tromp M. C., Van Boom J. H., Benne R. The sequence of the gene for cytochrome c oxidase subunit I, a frameshift containing gene for cytochrome c oxidase subunit II and seven unassigned reading frames in Trypanosoma brucei mitochondrial maxi-circle DNA. Nucleic Acids Res. 1984 Oct 11;12(19):7327–7344. doi: 10.1093/nar/12.19.7327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hunter W. N., Brown T., Anand N. N., Kennard O. Structure of an adenine-cytosine base pair in DNA and its implications for mismatch repair. Nature. 1986 Apr 10;320(6062):552–555. doi: 10.1038/320552a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Mahendran R., Spottswood M. R., Miller D. L. RNA editing by cytidine insertion in mitochondria of Physarum polycephalum. Nature. 1991 Jan 31;349(6308):434–438. doi: 10.1038/349434a0. [DOI] [PubMed] [Google Scholar]
  24. Maizels N., Weiner A. RNA editing. In search of a template. Nature. 1988 Aug 11;334(6182):469–470. doi: 10.1038/334469a0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Powell L. M., Wallis S. C., Pease R. J., Edwards Y. H., Knott T. J., Scott J. A novel form of tissue-specific RNA processing produces apolipoprotein-B48 in intestine. Cell. 1987 Sep 11;50(6):831–840. doi: 10.1016/0092-8674(87)90510-1. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Schuster W., Wissinger B., Unseld M., Brennicke A. Transcripts of the NADH-dehydrogenase subunit 3 gene are differentially edited in Oenothera mitochondria. EMBO J. 1990 Jan;9(1):263–269. doi: 10.1002/j.1460-2075.1990.tb08104.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shaw J. M., Feagin J. E., Stuart K., Simpson L. Editing of kinetoplastid mitochondrial mRNAs by uridine addition and deletion generates conserved amino acid sequences and AUG initiation codons. Cell. 1988 May 6;53(3):401–411. doi: 10.1016/0092-8674(88)90160-2. [DOI] [PubMed] [Google Scholar]
  30. Simpson L. Kinetoplast DNA in trypanosomid flagellates. Int Rev Cytol. 1986;99:119–179. doi: 10.1016/s0074-7696(08)61426-6. [DOI] [PubMed] [Google Scholar]
  31. Simpson L., Shaw J. RNA editing and the mitochondrial cryptogenes of kinetoplastid protozoa. Cell. 1989 May 5;57(3):355–366. doi: 10.1016/0092-8674(89)90911-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sloof P., van den Burg J., Voogd A., Benne R. The nucleotide sequence of a 3.2 kb segment of mitochondrial maxicircle DNA from Crithidia fasciculata containing the gene for cytochrome oxidase subunit III, the N-terminal part of the apocytochrome b gene and a possible frameshift gene; further evidence for the use of unusual initiator triplets in trypanosome mitochondria. Nucleic Acids Res. 1987 Jan 12;15(1):51–65. doi: 10.1093/nar/15.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stuart K. Trypanosomatids: mitochondrial RNA editing. Exp Parasitol. 1989 May;68(4):486–490. doi: 10.1016/0014-4894(89)90135-5. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Sturm N. R., Simpson L. Partially edited mRNAs for cytochrome b and subunit III of cytochrome oxidase from Leishmania tarentolae mitochondria: RNA editing intermediates. Cell. 1990 Jun 1;61(5):871–878. doi: 10.1016/0092-8674(90)90197-m. [DOI] [PubMed] [Google Scholar]
  36. Thomas S. M., Lamb R. A., Paterson R. G. Two mRNAs that differ by two nontemplated nucleotides encode the amino coterminal proteins P and V of the paramyxovirus SV5. Cell. 1988 Sep 9;54(6):891–902. doi: 10.1016/S0092-8674(88)91285-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tullis R. H., Rubin H. Calcium protects DNase I from proteinase K: a new method for the removal of contaminating RNase from DNase I. Anal Biochem. 1980 Sep 1;107(1):260–264. doi: 10.1016/0003-2697(80)90519-9. [DOI] [PubMed] [Google Scholar]
  38. Van der Spek H., Speijer D., Arts G. J., Van den Burg J., Van Steeg H., Sloof P., Benne R. RNA editing in transcripts of the mitochondrial genes of the insect trypanosome Crithidia fasciculata. EMBO J. 1990 Jan;9(1):257–262. doi: 10.1002/j.1460-2075.1990.tb08103.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vidal S., Curran J., Kolakofsky D. A stuttering model for paramyxovirus P mRNA editing. EMBO J. 1990 Jun;9(6):2017–2022. doi: 10.1002/j.1460-2075.1990.tb08330.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Volloch V., Schweitzer B., Rits S. Uncoupling of the synthesis of edited and unedited COIII RNA in Trypanosoma brucei. Nature. 1990 Feb 1;343(6257):482–484. doi: 10.1038/343482a0. [DOI] [PubMed] [Google Scholar]
  41. Wyatt J. R., Puglisi J. D., Tinoco I., Jr RNA folding: pseudoknots, loops and bulges. Bioessays. 1989 Oct;11(4):100–106. doi: 10.1002/bies.950110406. [DOI] [PubMed] [Google Scholar]
  42. van der Spek H., Arts G. J., van den Burg J., Sloof P., Benne R. The nucleotide sequence of mitochondrial maxicircle genes of Crithidia fasciculata. Nucleic Acids Res. 1989 Jun 26;17(12):4876–4876. doi: 10.1093/nar/17.12.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. van der Spek H., van den Burg J., Croiset A., van den Broek M., Sloof P., Benne R. Transcripts from the frameshifted MURF3 gene from Crithidia fasciculata are edited by U insertion at multiple sites. EMBO J. 1988 Aug;7(8):2509–2514. doi: 10.1002/j.1460-2075.1988.tb03098.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES