Abstract
Intracisternal A-particle (IAP) sequences are endogenous retrovirus-like mobile elements, or retrotransposons, present at 1,000 copies in the mouse genome. These elements transpose in a replicative manner via an RNA intermediate and its reverse transcription, and their transposition should therefore be tightly controlled by their transcription level. To analyze the in vivo pattern of expression of these retrovirus-like elements, we constructed several independent transgenic mice with either a complete IAP element marked with an intron or with the IAP promoter, or long terminal repeat (LTR), alone controlling the expression of a lacZ reporter gene with a nuclear localization signal. For all transgenic lines analyzed, IAP expression as determined by reverse transcription-PCR analysis was found to be essentially restricted to the male germ line. Furthermore, in situ 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining of all organs disclosed specific beta-galactosidase-positive blue cells only within the testis, found as patches along the seminiferous tubules and often organized as assemblies of 2, 4, 8, or 16 cells. Histochemical analyses of tissues from 13.5-day-old embryos to adults demonstrated that this LTR activity is restricted to gonocytes and premeiotic undifferentiated spermatogonia. Finally, analysis of the methylation status of both transgenes and endogenous IAP LTRs demonstrated identical patterns, with methylation in somatic tissues and hypomethylation in the testis. Transgenic mice therefore reveal an intrinsic, highly restricted IAP expression which had escaped detection in previous global Northern (RNA) blot analyses and with possible strong biological relevance, as IAP activation specifically within the germ line might be a way to generate diversity at the evolutionary level without being deleterious to individuals.
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- Bellvé A. R., Cavicchia J. C., Millette C. F., O'Brien D. A., Bhatnagar Y. M., Dym M. Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization. J Cell Biol. 1977 Jul;74(1):68–85. doi: 10.1083/jcb.74.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bird A. P. CpG-rich islands and the function of DNA methylation. Nature. 1986 May 15;321(6067):209–213. doi: 10.1038/321209a0. [DOI] [PubMed] [Google Scholar]
- Bonnerot C., Rocancourt D., Briand P., Grimber G., Nicolas J. F. A beta-galactosidase hybrid protein targeted to nuclei as a marker for developmental studies. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6795–6799. doi: 10.1073/pnas.84.19.6795. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brinster R. L., Zimmermann J. W. Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11298–11302. doi: 10.1073/pnas.91.24.11298. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CLERMONT Y., PEREY B. Quantitative study of the cell population of the seminiferous tubules in immature rats. Am J Anat. 1957 Mar;100(2):241–267. doi: 10.1002/aja.1001000205. [DOI] [PubMed] [Google Scholar]
- 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]
- Choi Y. C., Chae C. B. DNA hypomethylation and germ cell-specific expression of testis-specific H2B histone gene. J Biol Chem. 1991 Oct 25;266(30):20504–20511. [PubMed] [Google Scholar]
- Crowther P. J., Doherty J. P., Linsenmeyer M. E., Williamson M. R., Woodcock D. M. Revised genomic consensus for the hypermethylated CpG island region of the human L1 transposon and integration sites of full length L1 elements from recombinant clones made using methylation-tolerant host strains. Nucleic Acids Res. 1991 May 11;19(9):2395–2401. doi: 10.1093/nar/19.9.2395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Rooij D. G. Regulation of the proliferation of spermatogonial stem cells. J Cell Sci Suppl. 1988;10:181–194. doi: 10.1242/jcs.1988.supplement_10.14. [DOI] [PubMed] [Google Scholar]
- Doerfler W. Patterns of DNA methylation--evolutionary vestiges of foreign DNA inactivation as a host defense mechanism. A proposal. Biol Chem Hoppe Seyler. 1991 Aug;372(8):557–564. [PubMed] [Google Scholar]
- Duhl D. M., Vrieling H., Miller K. A., Wolff G. L., Barsh G. S. Neomorphic agouti mutations in obese yellow mice. Nat Genet. 1994 Sep;8(1):59–65. doi: 10.1038/ng0994-59. [DOI] [PubMed] [Google Scholar]
- Dupressoir A., Puech A., Heidmann T. IAP retrotransposons in the mouse liver as reporters of ageing. Biochim Biophys Acta. 1995 Dec 27;1264(3):397–402. doi: 10.1016/0167-4781(95)00181-6. [DOI] [PubMed] [Google Scholar]
- Dührsen U., Stahl J., Gough N. M. In vivo transformation of factor-dependent hemopoietic cells: role of intracisternal A-particle transposition for growth factor gene activation. EMBO J. 1990 Apr;9(4):1087–1096. doi: 10.1002/j.1460-2075.1990.tb08214.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Falzon M., Fewell J. W., Kuff E. L. EBP-80, a transcription factor closely resembling the human autoantigen Ku, recognizes single- to double-strand transitions in DNA. J Biol Chem. 1993 May 15;268(14):10546–10552. [PubMed] [Google Scholar]
- Falzon M., Kuff E. L. Multiple protein-binding sites in an intracisternal A particle long terminal repeat. J Virol. 1988 Nov;62(11):4070–4077. doi: 10.1128/jvi.62.11.4070-4077.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feenstra A., Fewell J., Lueders K., Kuff E. In vitro methylation inhibits the promotor activity of a cloned intracisternal A-particle LTR. Nucleic Acids Res. 1986 May 27;14(10):4343–4352. doi: 10.1093/nar/14.10.4343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furano A. V., Robb S. M., Robb F. T. The structure of the regulatory region of the rat L1 (L1Rn, long interspersed repeated) DNA family of transposable elements. Nucleic Acids Res. 1988 Oct 11;16(19):9215–9231. doi: 10.1093/nar/16.19.9215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galien R., Mercier G., Garcette M., Emanoil-Ravier R. ras oncogene activates the intracisternal A particle long terminal repeat promoter through a c-AMP response element. Oncogene. 1991 May;6(5):849–855. [PubMed] [Google Scholar]
- Gardiner-Garden M., Frommer M. CpG islands in vertebrate genomes. J Mol Biol. 1987 Jul 20;196(2):261–282. doi: 10.1016/0022-2836(87)90689-9. [DOI] [PubMed] [Google Scholar]
- Heidmann O., Heidmann T. Retrotransposition of a mouse IAP sequence tagged with an indicator gene. Cell. 1991 Jan 11;64(1):159–170. doi: 10.1016/0092-8674(91)90217-m. [DOI] [PubMed] [Google Scholar]
- Heidmann T., Heidmann O., Nicolas J. F. An indicator gene to demonstrate intracellular transposition of defective retroviruses. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2219–2223. doi: 10.1073/pnas.85.7.2219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hojman-Montes de Oca F., Lasneret J., Dianoux L., Canivet M., Ravicovitch-Ravier R., Périès J. Regulation of intracisternal A particles in mouse teratocarcinoma cells: involvement of DNA methylation in transcriptional control. Biol Cell. 1984;52(3):199–204. doi: 10.1111/j.1768-322x.1985.tb00337.x. [DOI] [PubMed] [Google Scholar]
- Hsiao W. L., Gattoni-Celli S., Weinstein I. B. Effects of 5-azacytidine on expression of endogenous retrovirus-related sequences in C3H 10T1/2 cells. J Virol. 1986 Mar;57(3):1119–1126. doi: 10.1128/jvi.57.3.1119-1126.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iguchi-Ariga S. M., Schaffner W. CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation. Genes Dev. 1989 May;3(5):612–619. doi: 10.1101/gad.3.5.612. [DOI] [PubMed] [Google Scholar]
- Kluin P. M., de Rooij D. G. A comparison between the morphology and cell kinetics of gonocytes and adult type undifferentiated spermatogonia in the mouse. Int J Androl. 1981 Aug;4(4):475–493. doi: 10.1111/j.1365-2605.1981.tb00732.x. [DOI] [PubMed] [Google Scholar]
- Kuff E. L., Fewell J. W. Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability. Mol Cell Biol. 1985 Mar;5(3):474–483. doi: 10.1128/mcb.5.3.474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuff E. L. Intracisternal A particles in mouse neoplasia. Cancer Cells. 1990 Dec;2(12):398–400. [PubMed] [Google Scholar]
- Kuff E. L., Lueders K. K. The intracisternal A-particle gene family: structure and functional aspects. Adv Cancer Res. 1988;51:183–276. doi: 10.1016/s0065-230x(08)60223-7. [DOI] [PubMed] [Google Scholar]
- Lathe R., Vilotte J. L., Clark A. J. Plasmid and bacteriophage vectors for excision of intact inserts. Gene. 1987;57(2-3):193–201. doi: 10.1016/0378-1119(87)90122-3. [DOI] [PubMed] [Google Scholar]
- Leiter E. H., Fewell J. W., Kuff E. L. Glucose induces intracisternal type A retroviral gene transcription and translation in pancreatic beta cells. J Exp Med. 1986 Jan 1;163(1):87–100. doi: 10.1084/jem.163.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lueders K. K., Fewell J. W., Morozov V. E., Kuff E. L. Selective expression of intracisternal A-particle genes in established mouse plasmacytomas. Mol Cell Biol. 1993 Dec;13(12):7439–7446. doi: 10.1128/mcb.13.12.7439. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lueders K. K., Mietz J. A. Structural analysis of type II variants within the mouse intracisternal A-particle sequence family. Nucleic Acids Res. 1986 Feb 11;14(3):1495–1510. doi: 10.1093/nar/14.3.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClelland M., Nelson M. Effect of site-specific methylation on DNA modification methyltransferases and restriction endonucleases. Nucleic Acids Res. 1992 May 11;20 (Suppl):2145–2157. doi: 10.1093/nar/20.suppl.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDonald J. F. Evolution and consequences of transposable elements. Curr Opin Genet Dev. 1993 Dec;3(6):855–864. doi: 10.1016/0959-437x(93)90005-a. [DOI] [PubMed] [Google Scholar]
- McGowan R., Campbell R., Peterson A., Sapienza C. Cellular mosaicism in the methylation and expression of hemizygous loci in the mouse. Genes Dev. 1989 Nov;3(11):1669–1676. doi: 10.1101/gad.3.11.1669. [DOI] [PubMed] [Google Scholar]
- Michaud E. J., van Vugt M. J., Bultman S. J., Sweet H. O., Davisson M. T., Woychik R. P. Differential expression of a new dominant agouti allele (Aiapy) is correlated with methylation state and is influenced by parental lineage. Genes Dev. 1994 Jun 15;8(12):1463–1472. doi: 10.1101/gad.8.12.1463. [DOI] [PubMed] [Google Scholar]
- Mietz J. A., Fewell J. W., Kuff E. L. Selective activation of a discrete family of endogenous proviral elements in normal BALB/c lymphocytes. Mol Cell Biol. 1992 Jan;12(1):220–228. doi: 10.1128/mcb.12.1.220. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mietz J. A., Grossman Z., Lueders K. K., Kuff E. L. Nucleotide sequence of a complete mouse intracisternal A-particle genome: relationship to known aspects of particle assembly and function. J Virol. 1987 Oct;61(10):3020–3029. doi: 10.1128/jvi.61.10.3020-3029.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgan R. A., Huang R. C. Correlation of undermethylation of intracisternal A-particle genes with expression in murine plasmacytomas but not in NIH/3T3 embryo fibroblasts. Cancer Res. 1984 Nov;44(11):5234–5241. [PubMed] [Google Scholar]
- Nur I., Pascale E., Furano A. V. The left end of rat L1 (L1Rn, long interspersed repeated) DNA which is a CpG island can function as a promoter. Nucleic Acids Res. 1988 Oct 11;16(19):9233–9251. doi: 10.1093/nar/16.19.9233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- OAKBERG E. F. A description of spermiogenesis in the mouse and its use in analysis of the cycle of the seminiferous epithelium and germ cell renewal. Am J Anat. 1956 Nov;99(3):391–413. doi: 10.1002/aja.1000990303. [DOI] [PubMed] [Google Scholar]
- Pikó L., Hammons M. D., Taylor K. D. Amounts, synthesis, and some properties of intracisternal A particle-related RNA in early mouse embryos. Proc Natl Acad Sci U S A. 1984 Jan;81(2):488–492. doi: 10.1073/pnas.81.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanes J. R., Rubenstein J. L., Nicolas J. F. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 1986 Dec 1;5(12):3133–3142. doi: 10.1002/j.1460-2075.1986.tb04620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Satyamoorthy K., Park K., Atchison M. L., Howe C. C. The intracisternal A-particle upstream element interacts with transcription factor YY1 to activate transcription: pleiotropic effects of YY1 on distinct DNA promoter elements. Mol Cell Biol. 1993 Nov;13(11):6621–6628. doi: 10.1128/mcb.13.11.6621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shen-Ong G. L., Cole M. D. Differing populations of intracisternal A-particle genes in myeloma tumors and mouse subspecies. J Virol. 1982 May;42(2):411–421. doi: 10.1128/jvi.42.2.411-421.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trelogan S. A., Martin S. L. Tightly regulated, developmentally specific expression of the first open reading frame from LINE-1 during mouse embryogenesis. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1520–1524. doi: 10.1073/pnas.92.5.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wivel N. A., Smith G. H. Distribution of intracisternal A-particles in a variety of normal and neoplastic mouse tissues. Int J Cancer. 1971 Jan 15;7(1):167–175. doi: 10.1002/ijc.2910070119. [DOI] [PubMed] [Google Scholar]
- Woodcock D. M., Crowther P. J., Diver W. P., Graham M., Bateman C., Baker D. J., Smith S. S. RglB facilitated cloning of highly methylated eukaryotic DNA: the human L1 transposon, plant DNA, and DNA methylated in vitro with human DNA methyltransferase. Nucleic Acids Res. 1988 May 25;16(10):4465–4482. doi: 10.1093/nar/16.10.4465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ymer S., Tucker W. Q., Campbell H. D., Young I. G. Nucleotide sequence of the intracisternal A-particle genome inserted 5' to the interleukin-3 gene of the leukemia cell line WEHI-3B. Nucleic Acids Res. 1986 Jul 25;14(14):5901–5918. doi: 10.1093/nar/14.14.5901. [DOI] [PMC free article] [PubMed] [Google Scholar]