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. 1992 Nov 25;20(22):6007–6014. doi: 10.1093/nar/20.22.6007

Three distinct RNA sequence elements are required for efficient apolipoprotein B (apoB) RNA editing in vitro.

J W Backus 1, H C Smith 1
PMCID: PMC334467  PMID: 1461733

Abstract

Apolipoprotein B (apoB) mRNA is edited in rat liver and intestine to convert a CAA glutamine codon to a UAA translational stop codon by the direct conversion of cytidine to uridine at nucleotide 6666. We have proposed the 'mooring sequence' model for apoB RNA editing, in which editing complexes (editosomes) assemble on specific apoB mRNA flanking sequences to direct this site-specific editing event. One sequence element (approx. nts 6671-81, the presumed 'mooring sequence') has been previously identified as necessary for editing. We have identified two additional sequence elements which are necessary for efficient editing: (1) a 5' 'Regulator' region which modulates editing efficiency and (2) a 'Spacer' region between the editing site and the 3' mooring sequence, whose distance is critical for efficient editing. Utilizing this data, we have induced editing at a cryptic site and have defined a 22 nucleotide 'cassette' of specific apoB sequence which is sufficient to support wild-type levels of editing in vitro in a background of distal apoB RNA sequence.

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Selected References

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

  1. Backus J. W., Smith H. C. Apolipoprotein B mRNA sequences 3' of the editing site are necessary and sufficient for editing and editosome assembly. Nucleic Acids Res. 1991 Dec 25;19(24):6781–6786. doi: 10.1093/nar/19.24.6781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Boström K., Garcia Z., Poksay K. S., Johnson D. F., Lusis A. J., Innerarity T. L. Apolipoprotein B mRNA editing. Direct determination of the edited base and occurrence in non-apolipoprotein B-producing cell lines. J Biol Chem. 1990 Dec 25;265(36):22446–22452. [PubMed] [Google Scholar]
  4. 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]
  5. Chen S. H., Li X. X., Liao W. S., Wu J. H., Chan L. RNA editing of apolipoprotein B mRNA. Sequence specificity determined by in vitro coupled transcription editing. J Biol Chem. 1990 Apr 25;265(12):6811–6816. [PubMed] [Google Scholar]
  6. 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]
  7. Davies M. S., Wallis S. C., Driscoll D. M., Wynne J. K., Williams G. W., Powell L. M., Scott J. Sequence requirements for apolipoprotein B RNA editing in transfected rat hepatoma cells. J Biol Chem. 1989 Aug 15;264(23):13395–13398. [PubMed] [Google Scholar]
  8. Driscoll D. M., Wynne J. K., Wallis S. C., Scott J. An in vitro system for the editing of apolipoprotein B mRNA. Cell. 1989 Aug 11;58(3):519–525. doi: 10.1016/0092-8674(89)90432-7. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Harris M. E., Hajduk S. L. Kinetoplastid RNA editing: in vitro formation of cytochrome b gRNA-mRNA chimeras from synthetic substrate RNAs. Cell. 1992 Mar 20;68(6):1091–1099. doi: 10.1016/0092-8674(92)90080-v. [DOI] [PubMed] [Google Scholar]
  11. Lau P. P., Chen S. H., Wang J. C., Chan L. A 40 kilodalton rat liver nuclear protein binds specifically to apolipoprotein B mRNA around the RNA editing site. Nucleic Acids Res. 1990 Oct 11;18(19):5817–5821. doi: 10.1093/nar/18.19.5817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Mikaelian I., Sergeant A. A general and fast method to generate multiple site directed mutations. Nucleic Acids Res. 1992 Jan 25;20(2):376–376. doi: 10.1093/nar/20.2.376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Navaratnam N., Patel D., Shah R. R., Greeve J. C., Powell L. M., Knott T. J., Scott J. An additional editing site is present in apolipoprotein B mRNA. Nucleic Acids Res. 1991 Apr 25;19(8):1741–1744. doi: 10.1093/nar/19.8.1741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Shah R. R., Knott T. J., Legros J. E., Navaratnam N., Greeve J. C., Scott J. Sequence requirements for the editing of apolipoprotein B mRNA. J Biol Chem. 1991 Sep 5;266(25):16301–16304. [PubMed] [Google Scholar]
  17. Smith H. C., Kuo S. R., Backus J. W., Harris S. G., Sparks C. E., Sparks J. D. In vitro apolipoprotein B mRNA editing: identification of a 27S editing complex. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1489–1493. doi: 10.1073/pnas.88.4.1489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sommer B., Köhler M., Sprengel R., Seeburg P. H. RNA editing in brain controls a determinant of ion flow in glutamate-gated channels. Cell. 1991 Oct 4;67(1):11–19. doi: 10.1016/0092-8674(91)90568-j. [DOI] [PubMed] [Google Scholar]
  19. Vallette F., Mege E., Reiss A., Adesnik M. Construction of mutant and chimeric genes using the polymerase chain reaction. Nucleic Acids Res. 1989 Jan 25;17(2):723–733. doi: 10.1093/nar/17.2.723. [DOI] [PMC free article] [PubMed] [Google Scholar]

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