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:

Identification of an imprinting control region affecting the expression of all transcripts in the Gnas cluster

Nature Genetics volume 38, pages 350–355 (2006)Cite this article

Abstract

Genomic imprinting results in allele-specific silencing according to parental origin1. Silencing is brought about by imprinting control regions (ICRs) that are differentially marked in gametogenesis2. The group of imprinted transcripts in the mouse Gnas cluster (Nesp, Nespas, Gnasxl, Exon 1A and Gnas) provides a model for analyzing the mechanisms of imprint regulation. We previously identified an ICR that specifically regulates the tissue-specific imprinted expression of the Gnas gene3. Here we identify a second ICR at the Gnas cluster. We show that a paternally derived targeted deletion of the germline differentially methylated region (DMR) associated with the antisense Nespas transcript unexpectedly affects both the expression of all transcripts in the cluster and methylation of two DMRs. Our results establish that the Nespas DMR is the principal ICR at the Gnas cluster and functions bidirectionally as a switch for modulating expression of the antagonistically acting genes Gnasxl and Gnas. Uniquely, the Nespas DMR acts on the downstream ICR at exon 1A to regulate tissue-specific imprinting of the Gnas gene.

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

Access options

Buy this article

  • Purchase on SpringerLink
  • 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 Nespas DMR region and its effect on transcription and methylation of Nesp.
Figure 2: Gnasxl expression is reduced and Gnasxl promoter methylation is unaltered in mice carrying ΔNAS-DMR (KO).
Figure 3: Expression of Exon 1A and the Gnas gene after paternal and maternal transmission of ΔNAS-DMR (KO).
Figure 4: The Exon 1A DMR is partially methylated on the paternal allele when ΔNAS-DMR is paternally inherited.
Figure 5: Summary of the transcriptional and methylation status of the Gnas cluster.

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

    Article  CAS  Google Scholar 

  3. Williamson, C.M. et al. A cis-acting control region is required exclusively for the tissue-specific imprinting of Gnas. Nat. Genet. 36, 894–899 (2004).

    Article  CAS  Google Scholar 

  4. Weinstein, L.S., Liu, J., Sakamoto, A., Xie, T. & Chen, M. Minireview: GNAS: normal and abnormal functions. Endocrinology 145, 5459–5464 (2004).

    Article  CAS  Google Scholar 

  5. 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 

  6. Kelsey, G. et al. Identification of imprinted loci by methylation-sensitive representational difference analysis: application to mouse distal chromosome 2. Genomics 62, 129–138 (1999).

    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. Ischia, R. et al. Molecular cloning and characterization of NESP55, a novel chromogranin-like precursor of a peptide with 5-HT1B receptor antagonist activity. J. Biol. Chem. 272, 11657–11662 (1997).

    Article  CAS  Google Scholar 

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

    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. 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 

  12. Abramowitz, J. et al. XLαs, the extra-long form of the α-subunit of the Gs G protein, is significantly longer than suspected, and so is its companion Alex. Proc. Natl. Acad. Sci. USA 101, 8366–8371 (2004).

    Article  CAS  Google Scholar 

  13. Williamson, C.M., Skinner, J.A., Kelsey, G. & Peters, J. Alternative non-coding splice variants of Nespas, an imprinted gene antisense to Nesp in the Gnas imprinting cluster. Mamm. Genome 13, 74–79 (2002).

    Article  CAS  Google Scholar 

  14. 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 

  15. Liu, J. et al. Identification of the control region for tissue-specific imprinting of the stimulatory G protein α-subunit. Proc. Natl. Acad. Sci. USA 102, 5513–5518 (2005).

    Article  CAS  Google Scholar 

  16. Wutz, A. et al. Imprinted expression of the Igf2r gene depends on an intronic CpG island. Nature 389, 745–749 (1997).

    Article  CAS  Google Scholar 

  17. 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 

  18. 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 

  19. Judson, H., Hayward, B.E., Sheridan, E. & Bonthron, D.T. A global disorder of imprinting in the human female germ line. Nature 416, 539–542 (2002).

    Article  CAS  Google Scholar 

  20. Liu, J., Nealon, J.G. & Weinstein, L.S. Distinct patterns of abnormal GNAS imprinting in familial and sporadic pseudohypoparathyroidism type IB. Hum. Mol. Genet. 14, 95–102 (2005).

    Article  CAS  Google Scholar 

  21. Plagge, A. et al. The imprinted signalling protein XLαs is required for postnatal adaptation to feeding. Nat. Genet. 36, 818–826 (2004).

    Article  CAS  Google Scholar 

  22. 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 

  23. 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 

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

    Article  CAS  Google Scholar 

  25. Sleutels, F., Zwart, R. & Barlow, D.P. The non-coding Air RNA is required for silencing autosomal imprinted genes. Nature 415, 810–813 (2002).

    Article  CAS  Google Scholar 

  26. 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 

  27. Bastepe, M. et al. Deletion of the NESP55 differentially methylated region causes loss of maternal GNAS imprints and pseudohypoparathyroidism type Ib. Nat. Genet. 37, 25–27 (2004).

    Article  Google Scholar 

  28. Moore, T. & Haig, D. Genomic imprinting in mammalian development: a parental tug-of-war. Trends Genet. 7, 45–49 (1991).

    Article  CAS  Google Scholar 

  29. Rivera-Perez, J.A., Mallo, M., Gendron-Maguire, M., Gridley, T. & Behringer, R.R. Goosecoid is not an essential component of the mouse gastrula organizer but is required for craniofacial and rib development. Development 121, 3005–3012 (1995).

    CAS  PubMed  Google Scholar 

  30. Bock, C. et al. BiQ Analyser: visualisation and quality control for DNA methylation data from bisulphite sequencing. Bioinformatics 21, 4067–4068 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B.M. Cattanach for comments on the manuscript; D. Papworth for statistical analyses; E.P. Evans for karyotype analysis; J. Bowler for injection of blastocysts; the Harwell Transgenic Facility for technical support for the targeting; the GEMS core at MRC Harwell for sequencing and pyrosequencing; J. Harrison, M. Harrison, L. Jones and A. Cameron for animal husbandry; and A. Ford's group for imaging services. This work was supported by the UK Medical Research Council and by the UK Biotechnology and Biological Sciences Research Council.

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

  3. Christine M Williamson and Martin D Turner: These authors contributed equally to this work.

Authors and Affiliations

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

    Christine M Williamson, Martin D Turner, Simon T Ball, Wade T Nottingham, Peter Glenister, Martin Fray, Zuzanna Tymowska-Lalanne, Nicola Powles-Glover, Mark Maconochie & Jo Peters

  2. The Babraham Institute, Cambridge, CB2 4AT, UK

    Antonius Plagge & Gavin Kelsey

  3. Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford, OX3 9DS, UK

    Mark Maconochie

  4. School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9QG, UK

    Wade T Nottingham

Authors
  1. Christine M Williamson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Martin D Turner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simon T Ball
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wade T Nottingham
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter Glenister
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Martin Fray
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zuzanna Tymowska-Lalanne
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Antonius Plagge
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nicola Powles-Glover
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Gavin Kelsey
    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.

Supplementary information

Supplementary Fig. 1

Sense and antisense promoter activity detected in the Nespas-Gnasxl region of the mouse Gnas imprinting cluster in a cell transfection assay. (PDF 269 kb)

Supplementary Fig. 2

Maternal inheritance of ΔNAS-DMR does not alter the imprinted expression of Nespas, Nesp, Gnasxl or Exon 1A. (PDF 439 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Williamson, C., Turner, M., Ball, S. et al. Identification of an imprinting control region affecting the expression of all transcripts in the Gnas cluster. Nat Genet 38, 350–355 (2006). https://doi.org/10.1038/ng1731

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1731

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