Skip to main content

Advertisement

SpringerLink
Log in

No evidence for cumulative effects in a Dnmt3b hypomorph across multiple generations

  • Published:
Mammalian Genome Aims and scope Submit manuscript

Abstract

Observations of inherited phenotypes that cannot be explained solely through genetic inheritance are increasing. Evidence points to transmission of non-DNA molecules in the gamete as mediators of the phenotypes. However, in most cases it is unclear what the molecules are, with DNA methylation, chromatin proteins, and small RNAs being the most prominent candidates. From a screen to generate novel mouse mutants of genes involved in epigenetic reprogramming, we produced a DNA methyltransferase 3b allele that is missing exon 13. Mice that are homozygous for the mutant allele have smaller stature and reduced viability, with particularly high levels of female post-natal death. Reduced DNA methylation was also detected at telocentric repeats and the X-linked Hprt gene. However, none of the abnormal phenotypes or DNA methylation changes worsened with multiple generations of homozygous mutant inbreeding. This suggests that in our model the abnormalities are reset each generation and the processes of transgenerational epigenetic reprogramming are effective in preventing their inheritance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Argentaro A, Yang JC, Chapman L, Kowalczyk MS, Gibbons RJ, Higgs DR, Neuhaus D, Rhodes D (2007) Structural consequences of disease-causing mutations in the ATRX-DNMT3-DNMT3L (ADD) domain of the chromatin-associated protein ATRX. Proc Natl Acad Sci USA 104:11939–11944

    Article  PubMed  CAS  Google Scholar 

  • Ashe A, Morgan DK, Whitelaw NC, Bruxner TJ, Vickaryous NK, Cox LL, Butterfield NC, Wicking C, Blewitt ME, Wilkins SJ, Anderson GJ, Cox TC, Whitelaw E (2008) A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development. Genome Biol 9:R182

    Article  PubMed  Google Scholar 

  • Biniszkiewicz D, Gribnau J, Ramsahoye B, Gaudet F, Eggan K, Humpherys D, Mastrangelo MA, Jun Z, Walter J, Jaenisch R (2002) Dnmt1 overexpression causes genomic hypermethylation, loss of imprinting, and embryonic lethality. Mol Cell Biol 22:2124–2135

    Article  PubMed  CAS  Google Scholar 

  • Blewitt ME, Vickaryous NK, Hemley SJ, Ashe A, Bruxner TJ, Preis JI, Arkell R, Whitelaw E (2005) An N-ethyl-N-nitrosourea screen for genes involved in variegation in the mouse. Proc Natl Acad Sci USA 102:7629–7634

    Article  PubMed  CAS  Google Scholar 

  • Bock C, Reither S, Mikeska T, Paulsen M, Walter J, Lengauer T (2005) BiQ Analyzer: visualization and quality control for DNA methylation data from bisulfite sequencing. Bioinformatics 21:4067–4068

    Article  PubMed  CAS  Google Scholar 

  • Bohacek J, Gapp K, Saab BJ, Mansuy IM (2013) Transgenerational epigenetic effects on brain functions. Biol Psychiatry 73(4):313–320

    Article  PubMed  Google Scholar 

  • Bourc’his D, Miniou P, Jeanpierre M, Molina Gomes D, Dupont J, De Saint-Basile G, Maraschio P, Tiepolo L, Viegas-Pequignot E (1999) Abnormal methylation does not prevent X inactivation in ICF patients. Cytogenet Cell Genet 84:245–252

    Article  PubMed  Google Scholar 

  • Brykczynska U, Hisano M, Erkek S, Ramos L, Oakeley EJ, Roloff TC, Beisel C, Schubeler D, Stadler MB, Peters AH (2010) Repressive and active histone methylation mark distinct promoters in human and mouse spermatozoa. Nat Struct Mol Biol 17:679–687

    Article  PubMed  CAS  Google Scholar 

  • Chong S, Vickaryous N, Ashe A, Zamudio N, Youngson N, Hemley S, Stopka T, Skoultchi A, Matthews J, Scott HS, de Kretser D, O’Bryan M, Blewitt M, Whitelaw E (2007) Modifiers of epigenetic reprogramming show paternal effects in the mouse. Nat Genet 39:614–622

    Article  PubMed  CAS  Google Scholar 

  • Danchin E, Charmantier A, Champagne FA, Mesoudi A, Pujol B, Blanchet S (2011) Beyond DNA: integrating inclusive inheritance into an extended theory of evolution. Nat Rev Genet 12:475–486

    Article  PubMed  CAS  Google Scholar 

  • Daxinger L, Whitelaw E (2012) Understanding transgenerational epigenetic inheritance via the gametes in mammals. Nat Rev Genet 13:153–162

    Article  PubMed  CAS  Google Scholar 

  • Daxinger L, Oey H, Apedaile A, Sutton J, Ashe A, Whitelaw E (2012) A forward genetic screen identifies eukaryotic translation initiation factor 3, subunit H (eIF3h), as an enhancer of variegation in the mouse. G3 (Bethesda) 2:1393–1396

    Article  CAS  Google Scholar 

  • Dhayalan A, Tamas R, Bock I, Tattermusch A, Dimitrova E, Kudithipudi S, Ragozin S, Jeltsch A (2011) The ATRX-ADD domain binds to H3 tail peptides and reads the combined methylation state of K4 and K9. Hum Mol Genet 20:2195–2203

    Article  PubMed  CAS  Google Scholar 

  • Gartler SM, Varadarajan KR, Luo P, Canfield TK, Traynor J, Francke U, Hansen RS (2004) Normal histone modifications on the inactive X chromosome in ICF and Rett syndrome cells: implications for methyl-CpG binding proteins. BMC Biol 2:21

    Article  PubMed  Google Scholar 

  • Gendrel AV, Apedaile A, Coker H, Termanis A, Zvetkova I, Godwin J, Tang YA, Huntley D, Montana G, Taylor S, Giannoulatou E, Heard E, Stancheva I, Brockdorff N (2012) Smchd1-dependent and -independent pathways determine developmental dynamics of CpG island methylation on the inactive X chromosome. Dev Cell 23:265–279

    Article  PubMed  CAS  Google Scholar 

  • Grandjean V, Gounon P, Wagner N, Martin L, Wagner KD, Bernex F, Cuzin F, Rassoulzadegan M (2009) The miR-124-Sox9 paramutation: RNA-mediated epigenetic control of embryonic and adult growth. Development 136:3647–3655

    Article  PubMed  CAS  Google Scholar 

  • Guenatri M, Bailly D, Maison C, Almouzni G (2004) Mouse centric and pericentric satellite repeats form distinct functional heterochromatin. J Cell Biol 166:493–505

    Article  PubMed  CAS  Google Scholar 

  • Guerrero-Bosagna C, Skinner MK (2012) Environmentally induced epigenetic transgenerational inheritance of phenotype and disease. Mol Cell Endocrinol 354:3–8

    Article  PubMed  CAS  Google Scholar 

  • Guerrero-Bosagna C, Settles M, Lucker B, Skinner MK (2010) Epigenetic transgenerational actions of vinclozolin on promoter regions of the sperm epigenome. PLoS One 5(9):e13100. doi:10.1371/journal.pone.0013100

  • Guibert S, Forne T, Weber M (2012) Global profiling of DNA methylation erasure in mouse primordial germ cells. Genome Res 22:633–641

    Article  PubMed  CAS  Google Scholar 

  • Hadchouel M, Farza H, Simon D, Tiollais P, Pourcel C (1987) Maternal inhibition of hepatitis B surface antigen gene expression in transgenic mice correlates with de novo methylation. Nature 329:454–456

    Article  PubMed  CAS  Google Scholar 

  • Hagleitner MM, Lankester A, Maraschio P, Hulten M, Fryns JP, Schuetz C, Gimelli G, Davies EG, Gennery A, Belohradsky BH, de Groot R, Gerritsen EJ, Mattina T, Howard PJ, Fasth A, Reisli I, Furthner D, Slatter MA, Cant AJ, Cazzola G, van Dijken PJ, van Deuren M, de Greef JC, van der Maarel SM, Weemaes CM (2008) Clinical spectrum of immunodeficiency, centromeric instability and facial dysmorphism (ICF syndrome). J Med Genet 45:93–99

    Article  PubMed  CAS  Google Scholar 

  • Hammoud SS, Nix DA, Zhang H, Purwar J, Carrell DT, Cairns BR (2009) Distinctive chromatin in human sperm packages genes for embryo development. Nature 460:473–478

    Google Scholar 

  • Hansen RS, Wijmenga C, Luo P, Stanek AM, Canfield TK, Weemaes CM, Gartler SM (1999) The DNMT3B DNA methyltransferase gene is mutated in the ICF immunodeficiency syndrome. Proc Natl Acad Sci USA 96:14412–14417

    Article  PubMed  CAS  Google Scholar 

  • Heyn H, Vidal E, Sayols S, Sanchez-Mut JV, Moran S, Medina I, Sandoval J, Simo-Riudalbas L, Szczesna K, Huertas D, Gatto S, Matarazzo MR, Dopazo J, Esteller M (2012) Whole-genome bisulfite DNA sequencing of a DNMT3B mutant patient. Epigenetics 7:542–550

    Article  PubMed  CAS  Google Scholar 

  • Jin B, Tao Q, Peng J, Soo HM, Wu W, Ying J, Fields CR, Delmas AL, Liu X, Qiu J, Robertson KD (2008) DNA methyltransferase 3B (DNMT3B) mutations in ICF syndrome lead to altered epigenetic modifications and aberrant expression of genes regulating development, neurogenesis and immune function. Hum Mol Genet 17:690–709

    Article  PubMed  CAS  Google Scholar 

  • Juriloff DM, Harris MJ (2012) Hypothesis: the female excess in cranial neural tube defects reflects an epigenetic drag of the inactivating X chromosome on the molecular mechanisms of neural fold elevation. Birth Defects Res A Clin Mol Teratol 94:849–855

    Article  PubMed  CAS  Google Scholar 

  • Kalitsis P, Griffiths B, Choo KH (2006) Mouse telocentric sequences reveal a high rate of homogenization and possible role in Robertsonian translocation. Proc Natl Acad Sci USA 103:8786–8791

    Article  PubMed  CAS  Google Scholar 

  • Kaneda M, Okano M, Hata K, Sado T, Tsujimoto N, Li E, Sasaki H (2004) Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting. Nature 429:900–903

    Article  PubMed  CAS  Google Scholar 

  • Li E, Bestor TH, Jaenisch R (1992) Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69:915–926

    Article  PubMed  CAS  Google Scholar 

  • Li JY, Lees-Murdock DJ, Xu GL, Walsh CP (2004) Timing of establishment of paternal methylation imprints in the mouse. Genomics 84:952–960

    Article  PubMed  CAS  Google Scholar 

  • Liu WM, Pang RT, Chiu PC, Wong BP, Lao K, Lee KF, Yeung WS (2012) Sperm-borne microRNA-34c is required for the first cleavage division in mouse. Proc Natl Acad Sci USA 109:490–494

    Article  PubMed  CAS  Google Scholar 

  • Morgan HD, Sutherland HG, Martin DI, Whitelaw E (1999) Epigenetic inheritance at the agouti locus in the mouse. Nat Genet 23:314–318

    Article  PubMed  CAS  Google Scholar 

  • Nelson VR, Nadeau JH (2010) Transgenerational genetic effects. Epigenomics 2:797–806

    Article  PubMed  CAS  Google Scholar 

  • Neuhaus IM, Beier DR (1998) Efficient localization of mutations by interval haplotype analysis. Mamm Genome 9(2):150–154

    Article  PubMed  CAS  Google Scholar 

  • Okano M, Bell DW, Haber DA, Li E (1999) DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 99:247–257

    Article  PubMed  CAS  Google Scholar 

  • Ooi SK, Qiu C, Bernstein E, Li K, Jia D, Yang Z, Erdjument-Bromage H, Tempst P, Lin SP, Allis CD, Cheng X, Bestor TH (2007) DNMT3L connects unmethylated lysine 4 of histone H3 to de novo methylation of DNA. Nature 448:714–717

    Article  PubMed  CAS  Google Scholar 

  • Otani J, Nankumo T, Arita K, Inamoto S, Ariyoshi M, Shirakawa M (2009) Structural basis for recognition of H3K4 methylation status by the DNA methyltransferase 3A ATRX-DNMT3-DNMT3L domain. EMBO Rep 10:1235–1241

    Article  PubMed  CAS  Google Scholar 

  • Preis JI, Downes M, Oates NA, Rasko JE, Whitelaw E (2003) Sensitive flow cytometric analysis reveals a novel type of parent-of-origin effect in the mouse genome. Curr Biol 13:955–959

    Article  PubMed  CAS  Google Scholar 

  • Rassoulzadegan M, Grandjean V, Gounon P, Vincent S, Gillot I, Cuzin F (2006) RNA-mediated non-Mendelian inheritance of an epigenetic change in the mouse. Nature 441:469–474

    Article  PubMed  CAS  Google Scholar 

  • Sutherland HG, Kearns M, Morgan HD, Headley AP, Morris C, Martin DI, Whitelaw E (2000) Reactivation of heritably silenced gene expression in mice. Mamm Genome 11:347–355

    Article  PubMed  CAS  Google Scholar 

  • Tada M, Tada T, Lefebvre L, Barton SC, Surani MA (1997) Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells. EMBO J 16:6510–6520

    Article  PubMed  CAS  Google Scholar 

  • Takada S, Paulsen M, Tevendale M, Tsai CE, Kelsey G, Cattanach BM, Ferguson-Smith AC (2002) Epigenetic analysis of the Dlk1-Gtl2 imprinted domain on mouse chromosome 12: implications for imprinting control from comparison with Igf2-H19. Hum Mol Genet 11:77–86

    Article  PubMed  CAS  Google Scholar 

  • Ueda Y, Okano M, Williams C, Chen T, Georgopoulos K, Li E (2006) Roles for Dnmt3b in mammalian development: a mouse model for the ICF syndrome. Development 133:1183–1192

    Article  PubMed  CAS  Google Scholar 

  • Velasco G, Hubé F, Rollin J, Neuillet D, Philippe C, Bouzinba-Segard H, Galvani A, Viegas-Péquignot E, Francastel C (2010) Dnmt3b recruitment through E2F6 transcriptional repressor mediates germ-line gene silencing in murine somatic tissues. Proc Natl Acad Sci USA 107(20):9281–9286

    Article  PubMed  Google Scholar 

  • Watanabe T, Tomizawa S, Mitsuya K, Totoki Y, Yamamoto Y, Kuramochi-Miyagawa S, Iida N, Hoki Y, Murphy PJ, Toyoda A, Gotoh K, Hiura H, Arima T, Fujiyama A, Sado T, Shibata T, Nakano T, Lin H, Ichiyanagi K, Soloway PD, Sasaki H (2011) Role for piRNAs and noncoding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus. Science 332:848–852

    Article  PubMed  CAS  Google Scholar 

  • Whitelaw NC, Chong S, Morgan DK, Nestor C, Bruxner TJ, Ashe A, Lambley E, Meehan R, Whitelaw E (2010) Reduced levels of two modifiers of epigenetic gene silencing, Dnmt3a and Trim28, cause increased phenotypic noise. Genome Biol 11:R111

    Article  PubMed  CAS  Google Scholar 

  • Xu GL, Bestor TH, Bourc’his D, Hsieh CL, Tommerup N, Bugge M, Hulten M, Qu X, Russo JJ, Viegas-Péquignot E (1999) Chromosome instability and immunodeficiency syndrome caused by mutations in a DNA methyltransferase gene. Nature 402(6758):187–191

    Article  PubMed  CAS  Google Scholar 

  • Youngson N (2012) Challenges and future prospects for non-genetic inheritance. Nongenet Inherit 1:1–8

    Article  Google Scholar 

  • Youngson NA, Whitelaw E (2008) Transgenerational epigenetic effects. Annu Rev Genomics Hum Genet 9:233–257

    Article  PubMed  CAS  Google Scholar 

  • Youngson NA, Vickaryous N, van der Horst A, Epp T, Harten S, Fleming JS, Khanna KK, de Kretser DM, Whitelaw E (2011) A missense mutation in the transcription factor Foxo3a causes teratomas and oocyte abnormalities in mice. Mamm Genome 22:235–248

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y, Jurkowska R, Soeroes S, Rajavelu A, Dhayalan A, Bock I, Rathert P, Brandt O, Reinhardt R, Fischle W, Jeltsch A (2010) Chromatin methylation activity of Dnmt3a and Dnmt3a/3L is guided by interaction of the ADD domain with the histone H3 tail. Nucleic Acids Res 38:4246–4253

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank En Li (China Novartis Institutes for BioMedical Research) for the Dnmt3b knockout mice. This study was supported by Australian NHMRC project grants to EW. ARR and KLL were supported by Australian Postgraduate awards. JMM is supported by an NHMRC Senior Research Fellowship. EW is supported by an NHMRC Australia Fellowship.

Author information

Author notes

  1. Emma Whitelaw

    Present address: School of Molecular Sciences, Department of Genetics, La Trobe Institute for Molecular Science, Melbourne, VIC, 3086, Australia

Authors and Affiliations

  1. Queensland Institute of Medical Research, Herston, Brisbane, QLD, 4006, Australia

    Neil A. Youngson, Trevor Epp, Amity R. Roberts, Lucia Daxinger, Alyson Ashe, Edward Huang, Sarah K. Harten, Graham F. Kay, Suyinn Chong & Emma Whitelaw

  2. Department of Pharmacology, School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia

    Neil A. Youngson

  3. Institute of Molecular Genetics of the ASCR, v. v. i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Trevor Epp

  4. School of Biomolecular and Physical Sciences, Griffith University, Nathan, QLD, 4111, Australia

    Amity R. Roberts

  5. School of Molecular Bioscience, University of Sydney, Sydney, NSW, 2006, Australia

    Krystal L. Lester & Jacqueline M. Matthews

  6. Division of Craniofacial Medicine, Department of Pediatrics, University of Washington, Seattle, WA, 98195, USA

    Timothy Cox

  7. Mater Research, TRI Building, Level 4, 37 Kent Street, Woolloongabba, QLD, 4102, Australia

    Suyinn Chong

Authors
  1. Neil A. Youngson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Trevor Epp
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Amity R. Roberts
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lucia Daxinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alyson Ashe
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Edward Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Krystal L. Lester
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sarah K. Harten
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Graham F. Kay
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Timothy Cox
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jacqueline M. Matthews
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Suyinn Chong
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Emma Whitelaw
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Emma Whitelaw.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 234 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Youngson, N.A., Epp, T., Roberts, A.R. et al. No evidence for cumulative effects in a Dnmt3b hypomorph across multiple generations. Mamm Genome 24, 206–217 (2013). https://doi.org/10.1007/s00335-013-9451-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-013-9451-5

Keywords

Search

Navigation