• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Oct 1990; 9(10): 3363–3368.
PMCID: PMC552074

Reverse transcriptase activity from human embryonal carcinoma cells NTera2D1.

Abstract

We have identified an RNA-dependent DNA polymerase activity in the microsomal fraction of human pluri-potential embryonal carcinoma cells NTera2D1, which are known to express the full length coding strand of the genomic Line-1 (L1) elements. This activity was classified as a reverse transcriptase (RT) based on its utilization of an RT specific synthetic poly(Cm) template in the presence of Mn2+ ions. Treatment of the cell by ultraviolet irradiation (200 erg/mm2) which resulted in a 2- to 3-fold enhancement of the RT activity, was required for the reproducible detection of the activity throughout the entire purification procedure. More than a 100-fold enrichment in RT activity was obtained by centrifugation in a glycerol step gradient and a linear sucrose density gradient followed by Sephacryl S-1000 gel filtration. These experiments demonstrated that the RT activity was associated with a macromolecular complex having the characteristics of a viral-like particle with a major protein component of 37 kd. The presence of L1 mRNA in RT-containing fractions suggests that the activity identified could originate from L1 elements and/or be involved in the mechanism of retroposition.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andrews PW. Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol. 1984 Jun;103(2):285–293. [PubMed]
  • Andrews PW, Damjanov I, Simon D, Banting GS, Carlin C, Dracopoli NC, Føgh J. Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. Lab Invest. 1984 Feb;50(2):147–162. [PubMed]
  • Bauer G, Hofschneider PH. An RNA-dependent DNA polymerase, different from the known viral reverse transcriptases, in the chicken system. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3025–3029. [PMC free article] [PubMed]
  • Chandra P, Steel LK. Purification, biochemical characterization and serological analysis of cellular deoxyribonucleic acid polymerases and a reverse transcriptase from spleen of a patient with myelofibrotic syndrome. Biochem J. 1977 Dec 1;167(3):513–524. [PMC free article] [PubMed]
  • Dalton AJ, Hellman A, Kalter SS, Helmke RJ. Ultrastructural comparison of placental virus with several type-c- oncogenic viruses. J Natl Cancer Inst. 1974 Apr;52(4):1379–1381. [PubMed]
  • Dirksen ER, Levy JA. Virus-like particles in placentas from normal individuals and patients with systemic lupus erythematosus. J Natl Cancer Inst. 1977 Oct;59(4):1187–1192. [PubMed]
  • Fanning T, Singer M. The LINE-1 DNA sequences in four mammalian orders predict proteins that conserve homologies to retrovirus proteins. Nucleic Acids Res. 1987 Mar 11;15(5):2251–2260. [PMC free article] [PubMed]
  • Garfinkel DJ, Boeke JD, Fink GR. Ty element transposition: reverse transcriptase and virus-like particles. Cell. 1985 Sep;42(2):507–517. [PubMed]
  • Gerard GF, Rottman F, Green M. Poly(2'-O-methylcytidylate)-oligodeoxyguanylate as a template for the ribonucleic acid directed deoxyribonucleic acid polymerase in ribonucleic acid tumor virus particles and a specific probe for the ribonucleic acid directed enzyme in transformed murine cells. Biochemistry. 1974 Apr 9;13(8):1632–1641. [PubMed]
  • Goblet C, Prost E, Whalen RG. One-step amplification of transcripts in total RNA using the polymerase chain reaction. Nucleic Acids Res. 1989 Mar 11;17(5):2144–2144. [PMC free article] [PubMed]
  • Gorman CM, Rigby PW, Lane DP. Negative regulation of viral enhancers in undifferentiated embryonic stem cells. Cell. 1985 Sep;42(2):519–526. [PubMed]
  • Hattori M, Kuhara S, Takenaka O, Sakaki Y. L1 family of repetitive DNA sequences in primates may be derived from a sequence encoding a reverse transcriptase-related protein. Nature. 1986 Jun 5;321(6070):625–628. [PubMed]
  • Ikegami S, Taguchi T, Ohashi M, Oguro M, Nagano H, Mano Y. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature. 1978 Oct 5;275(5679):458–460. [PubMed]
  • Krokan H, Schaffer P, DePamphilis ML. Involvement of eucaryotic deoxyribonucleic acid polymerases alpha and gamma in the replication of cellular and viral deoxyribonucleic acid. Biochemistry. 1979 Oct 2;18(20):4431–4443. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Loh TP, Sievert LL, Scott RW. Proviral sequences that restrict retroviral expression in mouse embryonal carcinoma cells. Mol Cell Biol. 1987 Oct;7(10):3775–3784. [PMC free article] [PubMed]
  • Löwer R, Löwer J, Frank H, Harzmann R, Kurth R. Human teratocarcinomas cultured in vitro produce unique retrovirus-like viruses. J Gen Virol. 1984 May;65(Pt 5):887–898. [PubMed]
  • Löwer J, Wondrak EM, Kurth R. Genome analysis and reverse transcriptase activity of human teratocarcinoma-derived retroviruses. J Gen Virol. 1987 Nov;68(Pt 11):2807–2815. [PubMed]
  • Mellor J, Malim MH, Gull K, Tuite MF, McCready S, Dibbayawan T, Kingsman SM, Kingsman AJ. Reverse transcriptase activity and Ty RNA are associated with virus-like particles in yeast. Nature. 1985 Dec 12;318(6046):583–586. [PubMed]
  • Mondal H, Hofschneider PH. Demonstration of free reverse transcriptase in the nuclei of embryonic tissues of the Japanese quail. Biochem Biophys Res Commun. 1983 Oct 14;116(1):303–311. [PubMed]
  • Rogers JH. The origin and evolution of retroposons. Int Rev Cytol. 1985;93:187–279. [PubMed]
  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [PubMed]
  • Skowronski J, Singer MF. Expression of a cytoplasmic LINE-1 transcript is regulated in a human teratocarcinoma cell line. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6050–6054. [PMC free article] [PubMed]
  • Skowronski J, Fanning TG, Singer MF. Unit-length line-1 transcripts in human teratocarcinoma cells. Mol Cell Biol. 1988 Apr;8(4):1385–1397. [PMC free article] [PubMed]
  • Switzer RC, 3rd, Merril CR, Shifrin S. A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem. 1979 Sep 15;98(1):231–237. [PubMed]
  • Taketo M, Tanaka M. A cellular enhancer of retrovirus gene expression in embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3748–3752. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...