Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1989 Jan 25;17(2):691–698. doi: 10.1093/nar/17.2.691

Characterization of single base substitutions in edited apolipoprotein B transcripts.

G E Tennyson 1, C A Sabatos 1, T L Eggerman 1, H B Brewer Jr 1
PMCID: PMC331612  PMID: 2464797

Abstract

Mature RNA transcripts from a single eukaryotic gene may contain different nucleotide sequences, ranging from alternately spliced exons to transcripts from separate alleles differing by only one base. Our laboratory and others have recently reported another class of RNA sequence differences, occurring in transcripts from the single copy apolipoprotein B (apoB) gene. A unique RNA editing mechanism allows expression of the CAA glutamine codon encoded by the apoB gene at nucleotide 6666, or terminates translation by the introduction of a premature UAA translational stop codon. In this study, we used the polymerase chain reaction (PCR) to amplify and characterize edited apoB RNA transcripts differing by a single nucleotide. Amplification and sequence analysis from small quantities of total RNA will facilitate the study of RNA editing and transcription in general.

Full text

PDF
691

Images in this article

Selected References

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

  1. 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]
  2. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  3. Coleman R. A., Haynes E. B., Sand T. M., Davis R. A. Developmental coordinate expression of triacylglycerol and small molecular weight apoB synthesis and secretion by rat hepatocytes. J Lipid Res. 1988 Jan;29(1):33–42. [PubMed] [Google Scholar]
  4. Edge S. B., Hoeg J. M., Triche T., Schneider P. D., Brewer H. B., Jr Cultured human hepatocytes. Evidence for metabolism of low density lipoproteins by a pathway independent of the classical low density lipoprotein receptor. J Biol Chem. 1986 Mar 15;261(8):3800–3806. [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Higuchi K., Hospattankar A. V., Law S. W., Meglin N., Cortright J., Brewer H. B., Jr Human apolipoprotein B (apoB) mRNA: identification of two distinct apoB mRNAs, an mRNA with the apoB-100 sequence and an apoB mRNA containing a premature in-frame translational stop codon, in both liver and intestine. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1772–1776. doi: 10.1073/pnas.85.6.1772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hospattankar A. V., Higuchi K., Law S. W., Meglin N., Brewer H. B., Jr Identification of a novel in-frame translational stop codon in human intestine apoB mRNA. Biochem Biophys Res Commun. 1987 Oct 14;148(1):279–285. doi: 10.1016/0006-291x(87)91107-7. [DOI] [PubMed] [Google Scholar]
  9. Hughes T. E., Sasak W. V., Ordovas J. M., Forte T. M., Lamon-Fava S., Schaefer E. J. A novel cell line (Caco-2) for the study of intestinal lipoprotein synthesis. J Biol Chem. 1987 Mar 15;262(8):3762–3767. [PubMed] [Google Scholar]
  10. Kane J. P. Apolipoprotein B: structural and metabolic heterogeneity. Annu Rev Physiol. 1983;45:637–650. doi: 10.1146/annurev.ph.45.030183.003225. [DOI] [PubMed] [Google Scholar]
  11. Kawasaki E. S., Clark S. S., Coyne M. Y., Smith S. D., Champlin R., Witte O. N., McCormick F. P. Diagnosis of chronic myeloid and acute lymphocytic leukemias by detection of leukemia-specific mRNA sequences amplified in vitro. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5698–5702. doi: 10.1073/pnas.85.15.5698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kogan S. C., Doherty M., Gitschier J. An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. N Engl J Med. 1987 Oct 15;317(16):985–990. doi: 10.1056/NEJM198710153171603. [DOI] [PubMed] [Google Scholar]
  13. Lee C. C., Wu X. W., Gibbs R. A., Cook R. G., Muzny D. M., Caskey C. T. Generation of cDNA probes directed by amino acid sequence: cloning of urate oxidase. Science. 1988 Mar 11;239(4845):1288–1291. doi: 10.1126/science.3344434. [DOI] [PubMed] [Google Scholar]
  14. Lee D. M., Dashti N., Mok T. Apolipoprotein B-100 is the major form of this apolipoprotein secreted by human intestinal Caco-2 cells. Biochem Biophys Res Commun. 1988 Oct 14;156(1):581–587. doi: 10.1016/s0006-291x(88)80882-9. [DOI] [PubMed] [Google Scholar]
  15. Mitsuhashi T., Tennyson G. E., Nikodem V. M. Alternative splicing generates messages encoding rat c-erbA proteins that do not bind thyroid hormone. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5804–5808. doi: 10.1073/pnas.85.16.5804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mitsuhashi T., Tennyson G., Nikodem V. Nucleotide sequence of novel cDNAs generated by alternative splicing of a rat thyroid hormone receptor gene transcript. Nucleic Acids Res. 1988 Jun 24;16(12):5697–5697. doi: 10.1093/nar/16.12.5697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Sarkar G., Sommer S. S. RNA amplification with transcript sequencing (RAWTS). Nucleic Acids Res. 1988 Jun 10;16(11):5197–5197. doi: 10.1093/nar/16.11.5197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scharf S. J., Horn G. T., Erlich H. A. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science. 1986 Sep 5;233(4768):1076–1078. doi: 10.1126/science.3461561. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Sparks J. D., Sparks C. E., Bolognino M., Roncone A. M., Jackson T. K., Amatruda J. M. Effects of nonketotic streptozotocin diabetes on apolipoprotein B synthesis and secretion by primary cultures of rat hepatocytes. J Clin Invest. 1988 Jul;82(1):37–43. doi: 10.1172/JCI113597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stoflet E. S., Koeberl D. D., Sarkar G., Sommer S. S. Genomic amplification with transcript sequencing. Science. 1988 Jan 29;239(4839):491–494. doi: 10.1126/science.3340835. [DOI] [PubMed] [Google Scholar]
  24. Todd J. A., Bell J. I., McDevitt H. O. HLA-DQ beta gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature. 1987 Oct 15;329(6140):599–604. doi: 10.1038/329599a0. [DOI] [PubMed] [Google Scholar]
  25. Wong C., Dowling C. E., Saiki R. K., Higuchi R. G., Erlich H. A., Kazazian H. H., Jr Characterization of beta-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA. 1987 Nov 26-Dec 2Nature. 330(6146):384–386. doi: 10.1038/330384a0. [DOI] [PubMed] [Google Scholar]
  26. Wrischnik L. A., Higuchi R. G., Stoneking M., Erlich H. A., Arnheim N., Wilson A. C. Length mutations in human mitochondrial DNA: direct sequencing of enzymatically amplified DNA. Nucleic Acids Res. 1987 Jan 26;15(2):529–542. doi: 10.1093/nar/15.2.529. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES