• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of narLink to Publisher's site
Nucleic Acids Res. May 27, 1986; 14(10): 4343–4352.
PMCID: PMC339865

In vitro methylation inhibits the promotor activity of a cloned intracisternal A-particle LTR.


We studied the relation between LTR methylation and expression of the family of endogenous retrovirus-like elements related to mouse intracisternal A-particles (IAP). Comparative HpaII/MspI and HhaI restriction analysis of genomic DNA's showed that in cells and tissues with a low level of IAP gene expression, HpaII and HhaI sites within the 5' LTR were heavily methylated, while in cells abundantly expressing IAP's 20 to 30% of the 5' LTRs were demethylated at these sites. The effects of methylation on the promoter activity of a cloned IAP 5' LTR was studied directly, using the plasmid pMIA5' L-cat in which this LTR was linked to the chloramphenicol acetyl transferase (CAT) gene. In vitro methylation of three HhaI sites located between -137 and -205 bp from the RNA start site of this LTR completely inactivated the promoter activity of pMIA5' L-cat transfected into COS7 cells. Methylation of a HpaII site located 94 bp downstream from the RNA start site reduced the promoter activity by 75%. The results show that methylation at sites both upstream and downstream from the RNA start site profoundly effects the promoter activity of this LTR and suggest that methylation within the 5' LTR can serve to regulate IAP gene expression in vivo.

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 (1.0M), 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.
  • Lueders KK, Kuff EL. Sequences associated with intracisternal A particles are reiterated in the mouse genome. Cell. 1977 Dec;12(4):963–972. [PubMed]
  • Pikó L, Hammons MD, Taylor KD. 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. [PMC free article] [PubMed]
  • Kuff EL, Fewell JW. 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. [PMC free article] [PubMed]
  • Burt DW, Reith AD, Brammar WJ. A retroviral provirus closely associated with the Ren-2 gene of DBA/2 mice. Nucleic Acids Res. 1984 Nov 26;12(22):8579–8593. [PMC free article] [PubMed]
  • Rechavi G, Givol D, Canaani E. Activation of a cellular oncogene by DNA rearrangement: possible involvement of an IS-like element. Nature. 1982 Dec 16;300(5893):607–611. [PubMed]
  • Horowitz M, Luria S, Rechavi G, Givol D. Mechanism of activation of the mouse c-mos oncogene by the LTR of an intracisternal A-particle gene. EMBO J. 1984 Dec 1;3(12):2937–2941. [PMC free article] [PubMed]
  • Hawley RG, Shulman MJ, Hozumi N. Transposition of two different intracisternal A particle elements into an immunoglobulin kappa-chain gene. Mol Cell Biol. 1984 Dec;4(12):2565–2572. [PMC free article] [PubMed]
  • Kuff EL, Feenstra A, Lueders K, Smith L, Hawley R, Hozumi N, Shulman M. Intracisternal A-particle genes as movable elements in the mouse genome. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1992–1996. [PMC free article] [PubMed]
  • Leiter EH, Kuff EL. Intracisternal Type A particles in murine pancreatic B cells. Immunocytochemical demonstration of increased antigen (p73) in genetically diabetic mice. Am J Pathol. 1984 Jan;114(1):46–55. [PMC free article] [PubMed]
  • Razin A, Szyf M. DNA methylation patterns. Formation and function. Biochim Biophys Acta. 1984 Sep 10;782(4):331–342. [PubMed]
  • Bird AP. DNA methylation--how important in gene control? Nature. 1984 Feb 9;307(5951):503–504. [PubMed]
  • Jaenisch R, Jähner D. Methylation, expression and chromosomal position of genes in mammals. Biochim Biophys Acta. 1984 May 15;782(1):1–9. [PubMed]
  • Doerfler W. DNA methylation and gene activity. Annu Rev Biochem. 1983;52:93–124. [PubMed]
  • Hojman-Montes de Oca F, Dianoux L, Peries J, Emanoil-Ravicovitch R. Intracisternal A particles: RNA expression and DNA methylation in murine teratocarcinoma cell lines. J Virol. 1983 Apr;46(1):307–310. [PMC free article] [PubMed]
  • Mays-Hoopes LL, Brown A, Huang RC. Methylation and rearrangement of mouse intracisternal a particle genes in development, aging, and myeloma. Mol Cell Biol. 1983 Aug;3(8):1371–1380. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Lasneret J, Canivet M, Hojman-Montes de Oca F, Tobaly J, Emanoil-Ravicovitch R, Peries J. Activation of intracisternal a particles by 5-azacytidine in mouse Ki-BALB cell line. Virology. 1983 Jul 30;128(2):485–489. [PubMed]
  • Morgan RA, Huang RC. 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]
  • Busslinger M, Hurst J, Flavell RA. DNA methylation and the regulation of globin gene expression. Cell. 1983 Aug;34(1):197–206. [PubMed]
  • Lueders KK, Fewell JW, Kuff EL, Koch T. The long terminal repeat of an endogenous intracisternal A-particle gene functions as a promoter when introduced into eucaryotic cells by transfection. Mol Cell Biol. 1984 Oct;4(10):2128–2135. [PMC free article] [PubMed]
  • Lueders KK, Kuff EL. Intracisternal A-particle genes: identification in the genome of Mus musculus and comparison of multiple isolates from a mouse gene library. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3571–3575. [PMC free article] [PubMed]
  • Parker BA, Stark GR. Regulation of simian virus 40 transcription: sensitive analysis of the RNA species present early in infections by virus or viral DNA. J Virol. 1979 Aug;31(2):360–369. [PMC free article] [PubMed]
  • Kruczek I, Doerfler W. Expression of the chloramphenicol acetyltransferase gene in mammalian cells under the control of adenovirus type 12 promoters: effect of promoter methylation on gene expression. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7586–7590. [PMC free article] [PubMed]
  • Nordheim A, Rich A. Negatively supercoiled simian virus 40 DNA contains Z-DNA segments within transcriptional enhancer sequences. Nature. 1983 Jun 23;303(5919):674–679. [PubMed]
  • Laimins LA, Khoury G, Gorman C, Howard B, Gruss P. Host-specific activation of transcription by tandem repeats from simian virus 40 and Moloney murine sarcoma virus. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6453–6457. [PMC free article] [PubMed]
  • Derse D, Caradonna SJ, Casey JW. Bovine leukemia virus long terminal repeat: a cell type-specific promoter. Science. 1985 Jan 18;227(4684):317–320. [PubMed]
  • Sodroski J, Patarca R, Rosen C, Wong-Staal F, Haseltine W. Location of the trans-activating region on the genome of human T-cell lymphotropic virus type III. Science. 1985 Jul 5;229(4708):74–77. [PubMed]
  • Christy RJ, Brown AR, Gourlie BB, Huang RC. Nucleotide sequences of murine intracisternal A-particle gene LTRs have extensive variability within the R region. Nucleic Acids Res. 1985 Jan 11;13(1):289–302. [PMC free article] [PubMed]
  • Ono M, Ohishi H. Long terminal repeat sequences of intracisternal A particle genes in the Syrian hamster genome: identification of tRNAPhe as a putative primer tRNA. Nucleic Acids Res. 1983 Oct 25;11(20):7169–7179. [PMC free article] [PubMed]
  • Couez D, Deschamps J, Kettmann R, Stephens RM, Gilden RV, Burny A. Nucleotide sequence analysis of the long terminal repeat of integrated bovine leukemia provirus DNA and of adjacent viral and host sequences. J Virol. 1984 Feb;49(2):615–620. [PMC free article] [PubMed]
  • Seiki M, Hattori S, Hirayama Y, Yoshida M. Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3618–3622. [PMC free article] [PubMed]
  • Bensimhon M, Gabarro-Arpa J, Ehrlich R, Reiss C. Physical characteristics in eucaryotic promoters. Nucleic Acids Res. 1983 Jul 11;11(13):4521–4540. [PMC free article] [PubMed]
  • Keller JM, Alwine JC. Activation of the SV40 late promoter: direct effects of T antigen in the absence of viral DNA replication. Cell. 1984 Feb;36(2):381–389. [PubMed]

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


Related citations in PubMed

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...