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.

  • Original Article
  • Published:

Site-specific DNA methylation by a complex of PU.1 and Dnmt3a/b

Oncogene volume 25, pages 2477–2488 (2006)Cite this article

Abstract

The Ets transcription factor PU.1 is a hematopoietic master regulator essential for the development of myeloid and B-cell lineages. As we previously reported, PU.1 sometimes represses transcription on forming a complex with mSin3A–histone deacetyl transferase–MeCP2. Here, we show an interaction between PU.1 and DNA methyltransferases, DNA methyltransferase (Dnmt)3a and Dnmt3b (Dnmt3s). Glutathione-S-transferase pulldown assay revealed that PU.1 directly interacted with the ATRX domain of Dnmt3s through the ETS domain. Dnmt3s repressed the transcriptional activity of PU.1 on a reporter construct with trimerized PU.1-binding sites. The repression was recovered by addition of 5-aza-deoxycitidine, a DNA methyltransferase inhibitor, but not trichostatin A, a histone deacetylase inhibitor. Bisulfite sequence analysis revealed that several CpG sites in the promoter region neighboring the PU.1-binding sites were methylated when Dnmt3s were coexpressed with PU.1. We also showed that the CpG sites in the p16INK4A promoter were methylated by overexpression of PU.1 in NIH3T3 cells, accompanied by a downregulation of p16INK4A gene expression. These results suggest that PU.1 may downregulate its target genes through an epigenetic modification such as DNA methylation.

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
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

Abbreviations

GST:

glutathione-S-transferase

G418:

geneticin

Tris:

2-amino-2-hydroxymethyl-1,3-propanediol

DMEM:

Dulbecco's modified Eagle's minimum essential medium

HEPES:

2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid

MOPS:

3-morpholinopropanesulfonic acid

PVDF:

polyvinylidene fluoride

PMSF:

phenylmethylsulfonyl fluoride

EDTA:

ethylenediamine-N, N, N′, N′-tetraacetic acid

SDS:

sodium dodecylsulfate

DOC:

sodium deoxycholate

NP-40:

Nodiet P-40

5-AZAdC:

5-aza-deoxycitidine

TSA:

trichostatin A

HDAC:

histone deacetyl transferase

CBP:

CRE-binding protein

Dnmt:

DNA methyltransferase

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

References

  • Bachman KE, Rountree MR, Baylin SB . (2001). J Biol Chem 276: 32282–32287.

  • Bassuk AG, Leiden JM . (1995). Immunity 3: 223–237.

  • Bestor T, Laudano A, Mattaliano R, Ingram V . (1988). J Mol Biol 203: 971–983.

  • Bird AP, Wolffe AP . (1999). Cell 99: 451–454.

  • Croce LD, Raker VA, Corsaro M, Fazi F, Fanelli M, Fuks F et al. (2002). Science 295: 1079–1082.

  • Datta J, Ghoshal K, Sharma SM, Tajima S, Jacob ST . (2003). J Cell Biochem 88: 855–864.

  • Eisenbeis CF, Singh H, Storb U . (1995). Gene Dev 9: 1377–1387.

  • Fuks F, Burgers WA, Godin N, Kasai M, Kouzarides T . (2001). EMBO J 20: 2536–2544.

  • Hara E, Smith R, Parry D, Tahara H, Stone S, Peters G . (1996). Mol Cell Biol 16: 859–867.

  • Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB . (1996). Proc Natl Acad Sci USA 93: 9821–9826.

  • Hwang CK, Kim CS, Choi HS, McKercher SR, Loh HH . (2004). J Biol Chem 279: 19764–19774.

  • Jenuwein T, Allis CD . (2001). Science 293: 1074–1080.

  • Kihara-Negishi F, Yamamoto H, Suzuki M, Yamada T, Sakurai T, Tamura T et al. (2001). Oncogene 20: 6039–6047.

  • Klemsz MJ, McKercher SR, Celeda A, Maki RA . (1990). Cell 61: 13–124.

  • Konishi Y, Tominaga M, Watanabe Y, Imamura F, Goldfarb A, Maki R et al. (1999). Oncogene 18: 6795–6805.

  • Linderson Y, Eberhard D, Malin S, Johansson A, Busslinger M, Pettersson S . (2004). EMBO Rep 3: 291–296.

  • Majumder S, Ghoshal K, Datta J, Bai S, Dong X, Quan N et al. (2002). J Biol Chem 277: 16048–16058.

  • Moreau-Gachelin F . (1994). Biochim Biophys Acta 1198: 149–163.

  • Moreau-Gachelin F, Ray D, Mattei MG, Tambourin P, Tavitian A . (1989). Oncogene 4: 1449–1456.

  • Moreau-Gachelin F, Wending F, Molina T, Denis N, Titeux M, Grimber G et al. (1996). Mol Cell Biol 16: 2453–2463.

  • Muller BU, Pabst T, Osato M, Asou N, Johansen LM, Minden MD et al. (2002). Blood 100: 998–1007.

  • Nagulapalli S, Pongubala JM, Atchison ML . (1995). J Immunol 155: 4330–4338.

  • Ng HH, Bird A . (1999). Curr Opin Genet Dev 9: 158–163.

  • Oelgeschlager M, Nuchprayoon I, Luscher D, Friedman AD . (1996). Mol Cell Biol 16: 4717–4725.

  • Ohtani N, Zebedee Z, Huot TJ, Stinson JA, Sugimoto M, Ohashi Y et al. (2001). Nature 409: 1067–1070.

  • Oikawa T, Yamada T . (2003). Gene 303: 11–34.

  • Oikawa T, Yamada T, Kihara-Negishi F, Yamamoto H, Kondoh N, Hitomi Y et al. (1999). Cell Death Differ 6: 599–608.

  • Okano M, Xie S, Li E . (1998). Nat Genet 19: 219–220.

  • Pongubala JM, Atchison ML . (1997). Proc Natl Acad Sci USA 94: 127–132.

  • Rao G, Rekhtman N, Cheng G, Krasikov T, Skoultchi AI . (1997). Oncogene 14: 123–131.

  • Robertson KD . (2001). Oncogene 20: 3139–3155.

  • Robertson KD . (2002). Oncogene 21: 5361–5379.

  • Robertson KD, Ait-Si-Ali S, Yokochi T, Wade PA, Jones PL, Wolffe AP . (2000). Nat Genet 25: 338–342.

  • Rosenbauer F, Wagner K, Kutok JL, Iwasaki H, Le Beau MM, Okuno Y et al. (2004). Nat Genet 36: 624–630.

  • Rountree MR, Bachman KE, Baylin SB . (2000). Nat Genet 25: 269–277.

  • Singal R, Ginder GD . (1999). Blood 93: 4059–4070.

  • Suzuki M, Yamada T, Kihara-Negishi F, Sakurai T, Oikawa T . (2003). Oncogene 22: 8688–8698.

  • Tatematsu KI, Yamazaki T, Ishikawa F . (2000). Genes Cells 5: 677–688.

  • Tenen DG, Hromas R, Licht JD, Zhang DE . (1997). Blood 90: 498–519.

  • Vangala RK, Heiss-Neumann MS, Rangatia JS, Singh SM, Schoch C, Tenen DG et al. (2003). Blood 101: 270–277.

  • Walsh CP, Bestor TH . (1999). Genes Dev 13: 26–34.

  • Yamada T, Kondoh N, Matsumoto M, Yoshida M, Maekawa A, Oikawa T . (1997). Blood 89: 1383–1393.

  • Yamamoto H, Kihara-Negishi F, Yamada T, Hashimoto Y, Oikawa T . (1999). Oncogene 18: 1495–1501.

  • Zhang DE, Hetherington CJ, Chen HM, Tenen DG . (1994). Mol Cell Biol 14: 373–381.

Download references

Acknowledgements

We wish to thank Dr I Kitabayashi, National Cancer Center, Tokyo, Japan, for expression plasmids of mSin3A and HDAC1, and Dr C Jones, The Salk Institute, La Jolla CA, USA, for pTK-100-Luc reporter plasmid. We also thank Dr N Kondoh, National Defense Medical College, Saitama, Japan, for the helpful advice with respect to bisulfite sequencing analysis. This work was mainly supported by a Grant-in-Aid for Encouragement of Young Scientist (to MS) from the Ministry of Education, Science and Culture of Japan, and a Grant from the Nishi Foundation, Tokyo, Japan. Financial support for TO from Dr J Akiyama, OBGYN Akiyama Memorial Hospital, Hakodate, Dr K Watanabe, Watanabe Clinic, Shizuoka and Dr S Kurakata, Sankyo Ltd, Tokyo, Japan is also acknowledged.

Author information

Authors and Affiliations

  1. Department of Cell Genetics, Sasaki Institute, Tokyo, Japan

    M Suzuki, T Yamada, F Kihara-Negishi, T Sakurai & T Oikawa

  2. Research Institute for Biological Sciences, Tokyo University of Science, Chiba, Japan

    M Suzuki & N Hozumi

  3. Institute for Genome Research, University of Tokushima, Tokushima, Japan

    E Hara

  4. Beth Israel Hospital and Harvard Medical School, Louis Pasteur, MA, USA

    D G Tenen

Authors
  1. M Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F Kihara-Negishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T Sakurai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E Hara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D G Tenen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. N Hozumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. T Oikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T Oikawa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Suzuki, M., Yamada, T., Kihara-Negishi, F. et al. Site-specific DNA methylation by a complex of PU.1 and Dnmt3a/b. Oncogene 25, 2477–2488 (2006). https://doi.org/10.1038/sj.onc.1209272

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1209272

Keywords

This article is cited by

Search

Advanced search

Quick links