This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fujita, N.
Right arrow Articles by Nakao, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fujita, N.
Right arrow Articles by Nakao, M.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, July 2000, p. 5107-5118, Vol. 20, No. 14
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Mechanism of Transcriptional Regulation by Methyl-CpG Binding Protein MBD1

Naoyuki Fujita,1,2 Nobuya Shimotake,3 Izuru Ohki,3 Tsutomu Chiba,2 Hideyuki Saya,1 Masahiro Shirakawa,3 and Mitsuyoshi Nakao1,*

Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, Kumamoto 860-0811,1 Division of Gastroenterology, Department of Internal Medicine, Kyoto University Post-Graduate School of Medicine, Sakyo-ku, Kyoto 606-8507,2 and Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara 630-0101,3 Japan

Received 21 December 1999/Returned for modification 10 February 2000/Accepted 24 April 2000

MBD1 is a mammalian protein that binds symmetrically methylated CpG sequences and regulates gene expression in association with DNA methylation. This protein possesses a conserved sequence, named methyl-CpG binding domain (MBD), among a family of methyl-CpG binding proteins that mediate the biological consequences of the methylation. In addition, MBD1 has at least five isoforms due to alternative splicing events, resulting in the presence of CXXC1, CXXC2, and CXXC3 in MBD1 isoforms v1 (MBD1v1) and MBD1v2, and CXXC1 and CXXC2 in MBD1v3 and -v4. In the present study, we have investigated the significance of MBD, CXXC, and the C-terminal transcriptional repression domain (TRD) in MBD1. A bacterially expressed MBD binds efficiently to densely methylated rather than to sparsely methylated DNAs. In both methylation-deficient Drosophila melanogaster SL2 cells and mammalian CHO-K1 cells, MBD1v1 represses transcription preferentially from both unmethylated and sparsely methylated promoters, while MBD1v3 inhibits densely methylated but not unmethylated promoter activities. The CXXC3 sequence in MBD1v1 is responsible for the ability to bind unmethylated promoter. Furthermore, we have constructed mutant-type MBD1s in which the functionally important residues Arg22, Arg30, Asp32, Tyr34, Arg44, Ser45, and Tyr52 are changed to alanine to investigate the correlation between the structure and function of the MBD in MBD1. Excepting those for Ser45 and Tyr52, none of the recombinant MBD mutants bound to the densely methylated or unmethylated DNAs, and green fluorescent protein-fused MBD1 mutants did not localize properly in the nucleus. All the MBD1v1 and -v3 mutants lost the activity of methylation-dependent gene repression. Based on these findings we have concluded that MBD1 acts as a transcriptional regulator depending on the density of methyl-CpG pairs through the cooperation of MBD, CXXC, and TRD sequences.

* Corresponding author. Mailing address: Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan. Phone: 81-96-373-5118. Fax: 81-96-373-5120. E-mail: mnakao{at}gpo.kumamoto-u.ac.jp.

Molecular and Cellular Biology, July 2000, p. 5107-5118, Vol. 20, No. 14
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Pendino, F., Nguyen, E., Jonassen, I., Dysvik, B., Azouz, A., Lanotte, M., Segal-Bendirdjian, E., Lillehaug, J. R. (2009). Functional involvement of RINF, retinoid-inducible nuclear factor (CXXC5), in normal and tumoral human myelopoiesis. Blood 113: 3172-3181 [Abstract] [Full Text]  
  • Minoshima, M., Bando, T., Sasaki, S., Fujimoto, J., Sugiyama, H. (2008). Pyrrole-imidazole hairpin polyamides with high affinity at 5'-CGCG-3' DNA sequence; influence of cytosine methylation on binding. Nucleic Acids Res 36: 2889-2894 [Abstract] [Full Text]  
  • Sakamoto, Y., Watanabe, S., Ichimura, T., Kawasuji, M., Koseki, H., Baba, H., Nakao, M. (2007). Overlapping Roles of the Methylated DNA-binding Protein MBD1 and Polycomb Group Proteins in Transcriptional Repression of HOXA Genes and Heterochromatin Foci Formation. J. Biol. Chem. 282: 16391-16400 [Abstract] [Full Text]  
  • Fatemi, M., Wade, P. A. (2006). MBD family proteins: reading the epigenetic code.. J. Cell Sci. 119: 3033-3037 [Abstract] [Full Text]  
  • Villa, R., Morey, L., Raker, V. A., Buschbeck, M., Gutierrez, A., De Santis, F., Corsaro, M., Varas, F., Bossi, D., Minucci, S., Pelicci, P. G., Di Croce, L. (2006). The methyl-CpG binding protein MBD1 is required for PML-RAR{alpha} function. Proc. Natl. Acad. Sci. USA 103: 1400-1405 [Abstract] [Full Text]  
  • Brackertz, M., Gong, Z., Leers, J., Renkawitz, R. (2006). p66{alpha} and p66{beta} of the Mi-2/NuRD complex mediate MBD2 and histone interaction. Nucleic Acids Res 34: 397-406 [Abstract] [Full Text]  
  • Jang, J.-S., Lee, S. J., Choi, J. E., Cha, S. I., Lee, E. B., Park, T. I., Kim, C. H., Lee, W. K., Kam, S., Choi, J.-Y., Kang, Y. M., Park, R.-W., Kim, I.-S., Cho, Y. L., Jung, T. H., Han, S. B., Park, J. Y. (2005). Methyl-CpG Binding Domain 1 Gene Polymorphisms and Risk of Primary Lung Cancer. Cancer Epidemiol. Biomarkers Prev. 14: 2474-2480 [Abstract] [Full Text]  
  • Ichimura, T., Watanabe, S., Sakamoto, Y., Aoto, T., Fujita, N., Nakao, M. (2005). Transcriptional Repression and Heterochromatin Formation by MBD1 and MCAF/AM Family Proteins. J. Biol. Chem. 280: 13928-13935 [Abstract] [Full Text]  
  • Danam, R. P., Howell, S. R., Brent, T. P., Harris, L. C. (2005). Epigenetic regulation of O6-methylguanine-DNA methyltransferase gene expression by histone acetylation and methyl-CpG binding proteins. Molecular Cancer Therapeutics 4: 61-69 [Abstract] [Full Text]  
  • Ayton, P. M., Chen, E. H., Cleary, M. L. (2004). Binding to Nonmethylated CpG DNA Is Essential for Target Recognition, Transactivation, and Myeloid Transformation by an MLL Oncoprotein. Mol. Cell. Biol. 24: 10470-10478 [Abstract] [Full Text]  
  • Fujita, N., Kajita, M., Taysavang, P., Wade, P. A. (2004). Hormonal Regulation of Metastasis-Associated Protein 3 Transcription in Breast Cancer Cells. Mol. Endocrinol. 18: 2937-2949 [Abstract] [Full Text]  
  • Cornelie, S., Hoebeke, J., Schacht, A.-M., Bertin, B., Vicogne, J., Capron, M., Riveau, G. (2004). Direct Evidence that Toll-like Receptor 9 (TLR9) Functionally Binds Plasmid DNA by Specific Cytosine-phosphate-guanine Motif Recognition. J. Biol. Chem. 279: 15124-15129 [Abstract] [Full Text]  
  • Watanabe, S., Ichimura, T., Fujita, N., Tsuruzoe, S., Ohki, I., Shirakawa, M., Kawasuji, M., Nakao, M. (2003). Methylated DNA-binding domain 1 and methylpurine-DNA glycosylase link transcriptional repression and DNA repair in chromatin. Proc. Natl. Acad. Sci. USA 100: 12859-12864 [Abstract] [Full Text]  
  • Fujita, N., Watanabe, S., Ichimura, T., Tsuruzoe, S., Shinkai, Y., Tachibana, M., Chiba, T., Nakao, M. (2003). Methyl-CpG Binding Domain 1 (MBD1) Interacts with the Suv39h1-HP1 Heterochromatic Complex for DNA Methylation-based Transcriptional Repression. J. Biol. Chem. 278: 24132-24138 [Abstract] [Full Text]  
  • Zhao, X., Ueba, T., Christie, B. R., Barkho, B., McConnell, M. J., Nakashima, K., Lein, E. S., Eadie, B. D., Willhoite, A. R., Muotri, A. R., Summers, R. G., Chun, J., Lee, K.-F., Gage, F. H. (2003). Mice lacking methyl-CpG binding protein 1 have deficits in adult neurogenesis and hippocampal function. Proc. Natl. Acad. Sci. USA 100: 6777-6782 [Abstract] [Full Text]  
  • Reese, B. E., Bachman, K. E., Baylin, S. B., Rountree, M. R. (2003). The Methyl-CpG Binding Protein MBD1 Interacts with the p150 Subunit of Chromatin Assembly Factor 1. Mol. Cell. Biol. 23: 3226-3236 [Abstract] [Full Text]  
  • Fujita, N., Watanabe, S., Ichimura, T., Ohkuma, Y., Chiba, T., Saya, H., Nakao, M. (2003). MCAF Mediates MBD1-Dependent Transcriptional Repression. Mol. Cell. Biol. 23: 2834-2843 [Abstract] [Full Text]  
  • Tojo, M., Matsuzaki, K., Minami, T., Honda, Y., Yasuda, H., Chiba, T., Saya, H., Fujii-Kuriyama, Y., Nakao, M. (2002). The Aryl Hydrocarbon Receptor Nuclear Transporter Is Modulated by the SUMO-1 Conjugation System. J. Biol. Chem. 277: 46576-46585 [Abstract] [Full Text]  
  • Koizume, S., Tachibana, K., Sekiya, T., Hirohashi, S., Shiraishi, M. (2002). Heterogeneity in the modification and involvement of chromatin components of the CpG island of the silenced human CDH1 gene in cancer cells. Nucleic Acids Res 30: 4770-4780 [Abstract] [Full Text]  
  • Guo, Y., Pakneshan, P., Gladu, J., Slack, A., Szyf, M., Rabbani, S. A. (2002). Regulation of DNA Methylation in Human Breast Cancer. EFFECT ON THE UROKINASE-TYPE PLASMINOGEN ACTIVATOR GENE PRODUCTION AND TUMOR INVASION. J. Biol. Chem. 277: 41571-41579 [Abstract] [Full Text]  
  • Brackertz, M., Boeke, J., Zhang, R., Renkawitz, R. (2002). Two Highly Related p66 Proteins Comprise a New Family of Potent Transcriptional Repressors Interacting with MBD2 and MBD3. J. Biol. Chem. 277: 40958-40966 [Abstract] [Full Text]  
  • Saito, M., Ishikawa, F. (2002). The mCpG-binding Domain of Human MBD3 Does Not Bind to mCpG but Interacts with NuRD/Mi2 Components HDAC1 and MTA2. J. Biol. Chem. 277: 35434-35439 [Abstract] [Full Text]  
  • Ordway, J. M., Curran, T. (2002). Methylation Matters: Modeling a Manageable Genome. Cell Growth Differ. 13: 149-162 [Full Text]  
  • Birke, M., Schreiner, S., Garcia-Cuellar, M.-P., Mahr, K., Titgemeyer, F., Slany, R. K. (2002). The MT domain of the proto-oncoprotein MLL binds to CpG-containing DNA and discriminates against methylation. Nucleic Acids Res 30: 958-965 [Abstract] [Full Text]  
  • Kudo, S, Nomura, Y, Segawa, M, Fujita, N, Nakao, M, Hammer, S, Schanen, C, Terai, I, Tamura, M (2002). Functional characterisation of MeCP2 mutations found in male patients with X linked mental retardation. J. Med. Genet. 39: 132-136 [Full Text]  
  • Carlone, D. L., Skalnik, D. G. (2001). CpG Binding Protein Is Crucial for Early Embryonic Development. Mol. Cell. Biol. 21: 7601-7606 [Abstract] [Full Text]  
  • Sekimata, M., Takahashi, A., Murakami-Sekimata, A., Homma, Y. (2001). Involvement of a Novel Zinc Finger Protein, MIZF, in Transcriptional Repression by Interacting with a Methyl-CpG-binding Protein, MBD2. J. Biol. Chem. 276: 42632-42638 [Abstract] [Full Text]  
  • Yu, F., Zingler, N., Schumann, G., Stratling, W. H. (2001). Methyl-CpG-binding protein 2 represses LINE-1 expression and retrotransposition but not Alu transcription. Nucleic Acids Res 29: 4493-4501 [Abstract] [Full Text]  
  • Magdinier, F., Wolffe, A. P. (2001). Selective association of the methyl-CpG binding protein MBD2 with the silent p14/p16 locus in human neoplasia. Proc. Natl. Acad. Sci. USA 10.1073/pnas.101617298v1 [Abstract] [Full Text]  
  • Yamada, H., Vijayachandra, K., Penner, C., Glick, A. (2001). Increased Sensitivity of Transforming Growth Factor (TGF) beta 1 Null Cells to Alkylating Agents Reveals a Novel Link between TGFbeta Signaling and O6-Methylguanine Methyltransferase Promoter Hypermethylation. J. Biol. Chem. 276: 19052-19058 [Abstract] [Full Text]  
  • Lee, J.-H., Voo, K. S., Skalnik, D. G. (2001). Identification and Characterization of the DNA Binding Domain of CpG-binding Protein. J. Biol. Chem. 276: 44669-44676 [Abstract] [Full Text]  
  • Bauer, S., Kirschning, C. J., Hacker, H., Redecke, V., Hausmann, S., Akira, S., Wagner, H., Lipford, G. B. (2001). Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc. Natl. Acad. Sci. USA 98: 9237-9242 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»