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. 1993 Sep;13(9):5383–5392. doi: 10.1128/mcb.13.9.5383

Synchronous expression of LINE-1 RNA and protein in mouse embryonal carcinoma cells.

S L Martin 1, D Branciforte 1
PMCID: PMC360242  PMID: 8395003

Abstract

L1, or LINE-1, is a repetitive DNA family found in all mammalian genomes that have been examined. At least a few individual members of the L1 family are functional transposable elements. Expression of these active elements leads to new insertions of L1 into the genomic DNA by the process of retrotransposition. We have detected coexpression of full-length, sense-strand L1 RNA transcripts and L1-encoded protein in mouse embryonal carcinoma cell lines. Both of these L1 expression products are candidates for intermediates in the retrotransposition process. L1 protein is found in what appear to be cytoplasmic aggregates and is not localized to any known cytoplasmic organelles. The six embryonal carcinoma cell lines tested were chosen to represent commitment to different developmental pathways in early mouse embryogenesis. The only two cell lines that express L1 are unique among the six in that they have a strong predilection to differentiate into extraembryonic endoderm. This observation is consistent with L1 expression and transposition in primordial germ cells of the mouse. An important implication of these studies is that L1 expression may provide a new marker for use in determining the origin of primordial germ cells during mouse embryogenesis.

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Selected References

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

  1. 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]
  2. Darmon M., Bottenstein J., Sato G. Neural differentiation following culture of embryonal carcinoma cells in a serum-free defined medium. Dev Biol. 1981 Jul 30;85(2):463–473. doi: 10.1016/0012-1606(81)90277-3. [DOI] [PubMed] [Google Scholar]
  3. Deragon J. M., Sinnett D., Labuda D. Reverse transcriptase activity from human embryonal carcinoma cells NTera2D1. EMBO J. 1990 Oct;9(10):3363–3368. doi: 10.1002/j.1460-2075.1990.tb07537.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dombroski B. A., Mathias S. L., Nanthakumar E., Scott A. F., Kazazian H. H., Jr Isolation of an active human transposable element. Science. 1991 Dec 20;254(5039):1805–1808. doi: 10.1126/science.1662412. [DOI] [PubMed] [Google Scholar]
  5. Doolittle R. F., Feng D. F., Johnson M. S., McClure M. A. Origins and evolutionary relationships of retroviruses. Q Rev Biol. 1989 Mar;64(1):1–30. doi: 10.1086/416128. [DOI] [PubMed] [Google Scholar]
  6. Dudley J. P. Discrete high molecular weight RNA transcribed from the long interspersed repetitive element L1Md. Nucleic Acids Res. 1987 Mar 25;15(6):2581–2592. doi: 10.1093/nar/15.6.2581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans J. P., Palmiter R. D. Retrotransposition of a mouse L1 element. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8792–8795. doi: 10.1073/pnas.88.19.8792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Freedman R. Protein chemistry. Folding into the right shape. Nature. 1987 Sep 17;329(6136):196–197. doi: 10.1038/329196a0. [DOI] [PubMed] [Google Scholar]
  9. Ginsburg M., Snow M. H., McLaren A. Primordial germ cells in the mouse embryo during gastrulation. Development. 1990 Oct;110(2):521–528. doi: 10.1242/dev.110.2.521. [DOI] [PubMed] [Google Scholar]
  10. Griffiths G., Simons K., Warren G., Tokuyasu K. T. Immunoelectron microscopy using thin, frozen sections: application to studies of the intracellular transport of Semliki Forest virus spike glycoproteins. Methods Enzymol. 1983;96:466–485. doi: 10.1016/s0076-6879(83)96041-x. [DOI] [PubMed] [Google Scholar]
  11. Gunning P., Ponte P., Okayama H., Engel J., Blau H., Kedes L. Isolation and characterization of full-length cDNA clones for human alpha-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol. 1983 May;3(5):787–795. doi: 10.1128/mcb.3.5.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hardies S. C., Martin S. L., Voliva C. F., Hutchison C. A., 3rd, Edgell M. H. An analysis of replacement and synonymous changes in the rodent L1 repeat family. Mol Biol Evol. 1986 Mar;3(2):109–125. doi: 10.1093/oxfordjournals.molbev.a040386. [DOI] [PubMed] [Google Scholar]
  13. Ilves H., Kahre O., Speek M. Translation of the rat LINE bicistronic RNAs in vitro involves ribosomal reinitiation instead of frameshifting. Mol Cell Biol. 1992 Sep;12(9):4242–4248. doi: 10.1128/mcb.12.9.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jubier-Maurin V., Dod B. J., Bellis M., Piechaczyk M., Roizes G. Comparative study of the L1 family in the genus Mus. Possible role of retroposition and conversion events in its concerted evolution. J Mol Biol. 1985 Aug 20;184(4):547–564. doi: 10.1016/0022-2836(85)90302-x. [DOI] [PubMed] [Google Scholar]
  15. Kazazian H. H., Jr, Wong C., Youssoufian H., Scott A. F., Phillips D. G., Antonarakis S. E. Haemophilia A resulting from de novo insertion of L1 sequences represents a novel mechanism for mutation in man. Nature. 1988 Mar 10;332(6160):164–166. doi: 10.1038/332164a0. [DOI] [PubMed] [Google Scholar]
  16. Kornfeld S., Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol. 1989;5:483–525. doi: 10.1146/annurev.cb.05.110189.002411. [DOI] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Leibold D. M., Swergold G. D., Singer M. F., Thayer R. E., Dombroski B. A., Fanning T. G. Translation of LINE-1 DNA elements in vitro and in human cells. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6990–6994. doi: 10.1073/pnas.87.18.6990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Loeb D. D., Padgett R. W., Hardies S. C., Shehee W. R., Comer M. B., Edgell M. H., Hutchison C. A., 3rd The sequence of a large L1Md element reveals a tandemly repeated 5' end and several features found in retrotransposons. Mol Cell Biol. 1986 Jan;6(1):168–182. doi: 10.1128/mcb.6.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Luan D. D., Korman M. H., Jakubczak J. L., Eickbush T. H. Reverse transcription of R2Bm RNA is primed by a nick at the chromosomal target site: a mechanism for non-LTR retrotransposition. Cell. 1993 Feb 26;72(4):595–605. doi: 10.1016/0092-8674(93)90078-5. [DOI] [PubMed] [Google Scholar]
  21. Luzio J. P., Brake B., Banting G., Howell K. E., Braghetta P., Stanley K. K. Identification, sequencing and expression of an integral membrane protein of the trans-Golgi network (TGN38). Biochem J. 1990 Aug 15;270(1):97–102. doi: 10.1042/bj2700097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Martin S. L. LINEs. Curr Opin Genet Dev. 1991 Dec;1(4):505–508. doi: 10.1016/s0959-437x(05)80199-6. [DOI] [PubMed] [Google Scholar]
  23. Martin S. L. Ribonucleoprotein particles with LINE-1 RNA in mouse embryonal carcinoma cells. Mol Cell Biol. 1991 Sep;11(9):4804–4807. doi: 10.1128/mcb.11.9.4804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Martin S. L., Voliva C. F., Burton F. H., Edgell M. H., Hutchison C. A., 3rd A large interspersed repeat found in mouse DNA contains a long open reading frame that evolves as if it encodes a protein. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2308–2312. doi: 10.1073/pnas.81.8.2308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Martin S. L., Voliva C. F., Hardies S. C., Edgell M. H., Hutchison C. A., 3rd Tempo and mode of concerted evolution in the L1 repeat family of mice. Mol Biol Evol. 1985 Mar;2(2):127–140. doi: 10.1093/oxfordjournals.molbev.a040340. [DOI] [PubMed] [Google Scholar]
  26. Mathias S. L., Scott A. F., Kazazian H. H., Jr, Boeke J. D., Gabriel A. Reverse transcriptase encoded by a human transposable element. Science. 1991 Dec 20;254(5039):1808–1810. doi: 10.1126/science.1722352. [DOI] [PubMed] [Google Scholar]
  27. Nagai K., Thøgersen H. C. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli. Methods Enzymol. 1987;153:461–481. doi: 10.1016/0076-6879(87)53072-5. [DOI] [PubMed] [Google Scholar]
  28. Nicolas J. F., Avner P., Gaillard J., Guenet J. L., Jakob H., Jacob F. Cell lines derived from teratocarcinomas. Cancer Res. 1976 Nov;36(11 Pt 2):4224–4231. [PubMed] [Google Scholar]
  29. Packer A. I., Manova K., Bachvarova R. F. A discrete LINE-1 transcript in mouse blastocysts. Dev Biol. 1993 May;157(1):281–283. doi: 10.1006/dbio.1993.1133. [DOI] [PubMed] [Google Scholar]
  30. Parchment R. E., Gramzinski R. A., Pierce G. B. Neoplastic embryoid bodies of embryonal carcinoma C44 as a source of blastocele-like fluid. Differentiation. 1990 Mar;43(1):51–58. doi: 10.1111/j.1432-0436.1990.tb00429.x. [DOI] [PubMed] [Google Scholar]
  31. Pierce G. B., Arechaga J., Jones A., Lewellyn A., Wells R. S. The fate of embryonal-carcinoma cells in mouse blastocysts. Differentiation. 1987;33(3):247–253. doi: 10.1111/j.1432-0436.1987.tb01564.x. [DOI] [PubMed] [Google Scholar]
  32. Pruitt S. C., Natoli T. A. Inhibition of differentiation by leukemia inhibitory factor distinguishes two induction pathways in P19 embryonal carcinoma cells. Differentiation. 1992 May;50(1):57–65. doi: 10.1111/j.1432-0436.1992.tb00486.x. [DOI] [PubMed] [Google Scholar]
  33. Schichman S. A., Severynse D. M., Edgell M. H., Hutchison C. A., 3rd Strand-specific LINE-1 transcription in mouse F9 cells originates from the youngest phylogenetic subgroup of LINE-1 elements. J Mol Biol. 1992 Apr 5;224(3):559–574. doi: 10.1016/0022-2836(92)90544-t. [DOI] [PubMed] [Google Scholar]
  34. Segal-Bendirdjian E., Heidmann T. Evidence for a reverse transcription intermediate for a marked line transposon in tumoral rat cells. Biochem Biophys Res Commun. 1991 Dec 16;181(2):863–870. doi: 10.1016/0006-291x(91)91270-m. [DOI] [PubMed] [Google Scholar]
  35. Shehee W. R., Chao S. F., Loeb D. D., Comer M. B., Hutchison C. A., 3rd, Edgell M. H. Determination of a functional ancestral sequence and definition of the 5' end of A-type mouse L1 elements. J Mol Biol. 1987 Aug 20;196(4):757–767. doi: 10.1016/0022-2836(87)90402-5. [DOI] [PubMed] [Google Scholar]
  36. Skowronski J., Fanning T. G., Singer M. F. Unit-length line-1 transcripts in human teratocarcinoma cells. Mol Cell Biol. 1988 Apr;8(4):1385–1397. doi: 10.1128/mcb.8.4.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Skowronski J., Singer M. F. 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. doi: 10.1073/pnas.82.18.6050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  39. Tooze J., Kern H. F., Fuller S. D., Howell K. E. Condensation-sorting events in the rough endoplasmic reticulum of exocrine pancreatic cells. J Cell Biol. 1989 Jul;109(1):35–50. doi: 10.1083/jcb.109.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Xiong Y., Eickbush T. H. Origin and evolution of retroelements based upon their reverse transcriptase sequences. EMBO J. 1990 Oct;9(10):3353–3362. doi: 10.1002/j.1460-2075.1990.tb07536.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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