Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

A cis-acting control region is required exclusively for the tissue-specific imprinting of Gnas

Nature Genetics volume 36, pages 894–899 (2004)Cite this article

Abstract

Genomic imprinting brings about allele-specific silencing according to parental origin1. Silencing is controlled by cis-acting regulatory regions that are differentially marked during gametogenesis and can act over hundreds of kilobases to silence many genes2,3,4,5,6. Two candidate imprinting control regions (ICRs) have been identified at the compact imprinted Gnas cluster on distal mouse chromosome 2, one at exon 1A upstream of Gnas itself7 and one covering the promoters for Gnasxl and the antisense Nespas (ref. 8). This imprinted cluster is complex, containing biallelic, maternally and paternally expressed transcripts that share exons9. Gnas itself is mainly biallelically expressed but is weakly paternally repressed in specific tissues10. Here we show that a paternally derived targeted deletion of the germline differentially methylated region at exon 1A abolishes tissue-specific imprinting of Gnas. This rescues the abnormal phenotype of mice with a maternally derived Gnas mutation11,12. Imprinting of alternative transcripts, Nesp, Gnasxl and Nespas (ref. 13), in the cluster is unaffected. The results establish that the differentially methylated region at exon 1A contains an imprinting control element that specifically regulates Gnas and comprises a characterized ICR for a gene that is only weakly imprinted in a minority of tissues. There must be a second ICR regulating the alternative transcripts.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Gene targeting of the exon 1A DMR.
Figure 2: Expression of transcripts as a consequence of the exon 1A DMR deletion.
Figure 3: +/ΔEx1A-DMR Oed-Sml/+, Oed-Sml/+ and wild-type mice at birth.
Figure 4: Summary of transcription and methylation status of the Gnas cluster in brown fat of wild-type mice (a) and in mice with paternal transmission of the exon 1A DMR deletion (b).

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Cattanach, B.M. & Kirk, M. Differential activity of maternally and paternally derived chromosome regions in mice. Nature 315, 496–498 (1985).

    Article  CAS  Google Scholar 

  2. Leighton, P.A., Ingram, R.S., Eggenschwiler, J., Efstratiadis, A. & Tilghman, S.M. Disruption of imprinting caused by deletion of the H19 gene region in mice. Nature 375, 34–39 (1995).

    Article  CAS  Google Scholar 

  3. Nicholls, R.D. & Knepper, J.L. Genome organisation, function and imprinting in Prader-Willi and Angelman syndromes. Ann. Rev. Genomics Hum. Genet. 2, 153–175 (2001).

    Article  CAS  Google Scholar 

  4. Zwart, R., Sleutels, F., Wutz, A., Schinkel, A.H. & Barlow, D.P. Bidirectional action of the Igf2r imprint control element on upstream and downstream imprinted genes. Genes Dev. 15, 2361–2366 (2001).

    Article  CAS  Google Scholar 

  5. Fitzpatrick, G.V., Soloway, P.D. & Higgins, M.J. Regional loss of imprinting and growth deficiency in mice with a targeted deletion of KvDMR1. Nat. Genet. 32, 426–431 (2002).

    Article  CAS  Google Scholar 

  6. Lin, S.-P. et al. Asymmetric regulation of imprinting on the maternal and paternal chromosomes at the Dlk1-Gtl2 imprinted cluster on mouse chromosome 12. Nat. Genet. 35, 97–102 (2003).

    Article  CAS  Google Scholar 

  7. Liu, J., Yu, S., Litman, D., Chen, W. & Weinstein, L.S. Identification of a methylation imprint mark within the mouse Gnas locus. Mol. Cell. Biol. 20, 5808–5817 (2000).

    Article  CAS  Google Scholar 

  8. Coombes, C. et al. Epigenetic properties and identification of an imprint mark in the Nesp-Gnasxl domain of the mouse Gnas imprinted locus. Mol. Cell. Biol. 23, 5475–5488 (2003).

    Article  CAS  Google Scholar 

  9. Peters, J. et al. A cluster of oppositely imprinted transcripts at the Gnas locus in the distal imprinting region of mouse chromosome 2. Proc. Natl. Acad. Sci. USA 96, 3830–3835 (1999).

    Article  CAS  Google Scholar 

  10. Yu, S. et al. Variable and tissue-specific hormone resistance in heterotrimeric Gs protein α-subunit (Gsα) knockout mice is due to tissue-specific imprinting of the Gsα gene. Proc. Natl. Acad. Sci. USA 95, 8715–8720 (1998).

    Article  CAS  Google Scholar 

  11. Cattanach, B.M., Peters, J., Ball, S. & Rasberry, C. Two imprinted gene mutations: three phenotypes. Hum. Mol. Genet. 9, 2263–2273 (2000).

    Article  CAS  Google Scholar 

  12. Skinner, J.A., Cattanach, B.M. & Peters, J. The imprinted oedematous-small mutation on mouse chromosome 2 identifies new roles for Gnas and Gnasxl in development. Genomics 80, 373–375 (2002).

    Article  CAS  Google Scholar 

  13. Wroe, S.F. et al. An imprinted transcript, antisense to Nesp, adds complexity to the cluster of imprinted genes at the mouse Gnas locus. Proc. Natl. Acad. Sci. USA 97, 3342–3346 (2000).

    Article  CAS  Google Scholar 

  14. Li, T. et al. Tissue-specific expression of antisense and sense transcripts at the imprinted Gnas locus. Genomics 69, 295–304 (2000).

    Article  CAS  Google Scholar 

  15. Chan, S.D.H. et al. Cloning and characterisation of the mouse Gsα promoter: activation by retinoic acid in F9 teratocarcinoma cells. Endocrine J. 2, 311–316 (1994).

    CAS  Google Scholar 

  16. Weinstein, L.S., Yu, S., Warner, D.R. & Liu, J. Endocrine manifestations of stimulatory G protein α-subunit mutations and the role of genomic imprinting. Endocrine Rev. 22, 675–705 (2001).

    CAS  Google Scholar 

  17. Liu, J. et al. A GNAS1 imprinting defect in pseudohypoparathyroidism type IB. J. Clin. Invest. 106, 1167–1174 (2000).

    Article  CAS  Google Scholar 

  18. Bastepe, M., Lane, A.H. & Juppner, H. Paternal uniparental isodisomy of chromosome 20q - and the resulting changes in GNAS1 methylation - as a plausible cause of pseudohypoparathyroidism. Am. J. Hum. Genet. 68, 1283–1289 (2001).

    Article  CAS  Google Scholar 

  19. Bastepe, M. et al. Autosomal dominant pseudohypoparathyroidism type Ib is associated with a heterozygous microdeletion that likely disrupts a putative imprinting control element of GNAS. J. Clin. Invest. 112, 1255–1263 (2003).

    Article  CAS  Google Scholar 

  20. Spahn, L. & Barlow, D.P. An ICE pattern crystallizes. Nat. Genet. 35, 11–12 (2003).

    Article  CAS  Google Scholar 

  21. Ischia, R. et al. Molecular cloning and characterisation of NESP55, a novel chromogranin-like precursor of a peptide with 5-HT1B receptor antagonist activity. J. Biol. Chem. 17, 11657–11662 (1997).

    Article  Google Scholar 

  22. Kehlenbach, R.H., Matthey, J. & Huttner, W.B. XL alpha s is a new type of G protein. Nature 372, 804–808 (1994).

    Article  CAS  Google Scholar 

  23. Hayward, B.E. et al. The human GNAS1 gene is imprinted and encodes distinct paternally and biallelically expressed G proteins. Proc. Natl. Acad. Sci. USA 95, 10038–10043 (1998).

    Article  CAS  Google Scholar 

  24. Hayward, B.E., Moran, V., Strain, L. & Bonthron, D.T. Bidirectional imprinting of a single gene: GNAS1 encodes maternally, paternally, and biallelically derived proteins. Proc. Natl. Acad. Sci. USA 95, 15475–15480 (1998).

    Article  CAS  Google Scholar 

  25. Hayward, B.E. & Bonthron, D.T. An imprinted antisense transcript at the human GNAS1 locus. Hum. Mol. Genet. 9, 835–841 (2000).

    Article  CAS  Google Scholar 

  26. Storck, T., Krüth, U., Kolhekar, R., Sprengel, R. & Seeburg, P.H. Rapid construction in yeast of complex targeting vectors for gene manipulation in the mouse. Nucleic Acids Res. 24, 4594–4596 (1996).

    Article  CAS  Google Scholar 

  27. Bunting, M., Bernstein, K.E., Greer, J.M., Capecchi, M.R. & Thomas, K.R. Targeting genes for self-excision in the germline. Genes Dev. 13, 1524–1528 (1999).

    Article  CAS  Google Scholar 

  28. Ramirez-Solis, R., Davis, A.C. & Bradley, A. Gene targeting in embryonic stem cells. Methods Enzymol. 225, 855–878 (1993).

    Article  CAS  Google Scholar 

  29. Hough, T. et al. Novel phenotypes identified by plasma biochemical screen in the mouse. Mamm. Genome 13, 595–602 (2002).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B.M. Cattanach and G. Kelsey for useful comments on the manuscript; P. Denny for advice and help in obtaining the sequence of the chromosome 2 mouse BAC clone RP23-439H2 from the UK Mouse Genome Sequencing Programme; J. Bowler for injection of blastocysts; the Harwell Transgenic Facility for technical support for the targeting; M. Harrison, L. Jones and J. McNaughton for animal husbandry; and A. Ford's group for Imaging Services. Other sequencing was carried out by the GEMS core at MRC Harwell.

Author information

Author notes

  1. Wade T Nottingham

    Present address: Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford, OX3 9DS, UK

  2. Mark Maconochie

    Present address: School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9QG, UK

Authors and Affiliations

  1. MRC Mammalian Genetics Unit, Harwell, Oxfordshire, OX11 0RD, UK

    Christine M Williamson, Simon T Ball, Wade T Nottingham, Judith A Skinner, Martin D Turner, Nicola Powles, Tertius Hough, David Papworth, Mark Maconochie & Jo Peters

  2. Developmental Genetics Programme, The Babraham Institute, Cambridge, CB2 4AT, UK

    Antonius Plagge

  3. Department of Clinical Chemistry, University of Liverpool, 4th Floor, Duncan Building, Daulby Street, Liverpool, L69 3GA, UK

    William D Fraser

Authors
  1. Christine M Williamson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Simon T Ball
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wade T Nottingham
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Judith A Skinner
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Antonius Plagge
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Martin D Turner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nicola Powles
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tertius Hough
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. David Papworth
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. William D Fraser
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Mark Maconochie
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jo Peters
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jo Peters.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Williamson, C., Ball, S., Nottingham, W. et al. A cis-acting control region is required exclusively for the tissue-specific imprinting of Gnas. Nat Genet 36, 894–899 (2004). https://doi.org/10.1038/ng1398

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1398

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing