Development, Vol 127, Issue 16 3419-3428, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Deletion of a silencer element disrupts H19 imprinting independently of a DNA methylation epigenetic switch

RA Drewell, JD Brenton, JF Ainscough, SC Barton, KJ Hilton, KL Arney, L Dandolo and MA Surani
Wellcome/CRC Institute of Cancer and Developmental Biology and Physiological Laboratory, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK.

The H19 imprinted gene is silenced when paternally inherited and active only when inherited maternally. This is thought to involve a cis-acting control region upstream of H19 that is responsible for regulating a number of functions including DNA methylation, asynchronous replication of parental chromosomes and an insulator. Here we report on the function of a 1.2 kb upstream element in the mouse, which was previously shown to function as a bi-directional silencer in Drosophila. The cre-loxP-mediated targeted deletion of the 1.2 kb region had no effect on the maternal allele. However, there was loss of silencing of the paternal allele in many endodermal and other tissues. The pattern of expression was very similar to the expression pattern conferred by the enhancer elements downstream of H19. We could not detect an effect on the expression of the neighbouring imprinted Igf2 gene, suggesting that the proposed boundary element insulating this gene from the downstream enhancers was unaffected. Despite derepression of the paternal H19 allele, the deletion surprisingly did not affect the differential DNA methylation of the locus, which displayed an appropriate epigenetic switch in the parental germlines. Furthermore, the characteristic asynchronous pattern of DNA replication at H19 was also not disrupted by the deletion, suggesting that the sequences that mediate this were also intact. The silencer is therefore part of a complex cis-regulatory region upstream of the H19 gene and acts specifically to ensure the repression of the paternal allele, without a predominant effect on the epigenetic switch in the germline.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				P. C. Haycock and M. Ramsay Exposure of Mouse Embryos to Ethanol During Preimplantation Development: Effect on DNA Methylation in the H19 Imprinting Control Region Biol Reprod, October 1, 2009; 81(4): 618 - 627. [Abstract] [Full Text] [PDF]

				Y. Chen, A. Dhupelia, and C. J. Schoenherr The Igf2/H19 imprinting control region exhibits sequence-specific and cell-type-dependent DNA methylation-mediated repression Nucleic Acids Res., February 1, 2009; 37(3): 793 - 803. [Abstract] [Full Text] [PDF]

				O. S. Akbari, E. Bae, H. Johnsen, A. Villaluz, D. Wong, and R. A. Drewell A novel promoter-tethering element regulates enhancer-driven gene expression at the bithorax complex in the Drosophila embryo Development, January 1, 2008; 135(1): 123 - 131. [Abstract] [Full Text] [PDF]

				N. Engel, J. L. Thorvaldsen, and M. S. Bartolomei CTCF binding sites promote transcription initiation and prevent DNA methylation on the maternal allele at the imprinted H19/Igf2 locus Hum. Mol. Genet., October 1, 2006; 15(19): 2945 - 2954. [Abstract] [Full Text] [PDF]

				J. L. Thorvaldsen, A. M. Fedoriw, S. Nguyen, and M. S. Bartolomei Developmental Profile of H19 Differentially Methylated Domain (DMD) Deletion Alleles Reveals Multiple Roles of the DMD in Regulating Allelic Expression and DNA Methylation at the Imprinted H19/Igf2 Locus Mol. Cell. Biol., February 15, 2006; 26(4): 1245 - 1258. [Abstract] [Full Text] [PDF]

				V. Pant, S. Kurukuti, E. Pugacheva, S. Shamsuddin, P. Mariano, R. Renkawitz, E. Klenova, V. Lobanenkov, and R. Ohlsson Mutation of a Single CTCF Target Site within the H19 Imprinting Control Region Leads to Loss of Igf2 Imprinting and Complex Patterns of De Novo Methylation upon Maternal Inheritance Mol. Cell. Biol., April 15, 2004; 24(8): 3497 - 3504. [Abstract] [Full Text] [PDF]

				M. Vernucci, F. Cerrato, P. V. Pedone, L. Dandolo, C. B. Bruni, and A. Riccio Developmentally regulated functions of the H19 differentially methylated domain Hum. Mol. Genet., February 1, 2004; 13(3): 353 - 361. [Abstract] [Full Text] [PDF]

				S. SCHOENFELDER and R. PARO Drosophila Su(Hw) Regulates an Evolutionarily Conserved Silencer from the Mouse H19 Imprinting Control Region Cold Spring Harb Symp Quant Biol, January 1, 2004; 69(0): 47 - 54. [Abstract] [PDF]

				P. E. Szabo, S.-H. E. Tang, M. R. Reed, F. J. Silva, W. M. K. Tsark, and J. R. Mann The chicken {beta}-globin insulator element conveys chromatin boundary activity but not imprinting at the mouse Igf2/H19 domain Development, March 4, 2003; 129(4): 897 - 904. [Abstract] [Full Text] [PDF]

				S. Lopes, A. Lewis, P. Hajkova, W. Dean, J. Oswald, T. Forne, A. Murrell, M. Constancia, M. Bartolomei, J. Walter, et al. Epigenetic modifications in an imprinting cluster are controlled by a hierarchy of DMRs suggesting long-range chromatin interactions Hum. Mol. Genet., February 1, 2003; 12(3): 295 - 305. [Abstract] [Full Text] [PDF]

				N. Hori, H. Nakano, T. Takeuchi, H. Kato, S. Hamaguchi, M. Oshimura, and K. Sato A Dyad Oct-binding Sequence Functions as a Maintenance Sequence for the Unmethylated State within the H19/Igf2-imprinted Control Region J. Biol. Chem., July 26, 2002; 277(31): 27960 - 27967. [Abstract] [Full Text] [PDF]

				J. L. Thorvaldsen, M. R. W. Mann, O. Nwoko, K. L. Duran, and M. S. Bartolomei Analysis of Sequence Upstream of the Endogenous H19 Gene Reveals Elements Both Essential and Dispensable for Imprinting Mol. Cell. Biol., April 15, 2002; 22(8): 2450 - 2462. [Abstract] [Full Text] [PDF]

				R. A. Drewell, C. J. Goddard, J. O. Thomas, and M. A. Surani Methylation-dependent silencing at the H19 imprinting control region by MeCP2 Nucleic Acids Res., March 1, 2002; 30(5): 1139 - 1144. [Abstract] [Full Text] [PDF]

				R. A. Drewell, K. L. Arney, T. Arima, S. C. Barton, J. D. Brenton, and M. A. Surani Novel conserved elements upstream of the H19 gene are transcribed and act as mesodermal enhancers Development, January 3, 2002; 129(5): 1205 - 1213. [Abstract] [Full Text] [PDF]

				L. S. Weinstein, S. Yu, D. R. Warner, and J. Liu Endocrine Manifestations of Stimulatory G Protein {alpha}-Subunit Mutations and the Role of Genomic Imprinting Endocr. Rev., October 1, 2001; 22(5): 675 - 705. [Abstract] [Full Text] [PDF]

				R. I. Gregory, T. E. Randall, C. A. Johnson, S. Khosla, I. Hatada, L. P. O'Neill, B. M. Turner, and R. Feil DNA Methylation Is Linked to Deacetylation of Histone H3, but Not H4, on the Imprinted Genes Snrpn and U2af1-rs1 Mol. Cell. Biol., August 15, 2001; 21(16): 5426 - 5436. [Abstract] [Full Text] [PDF]

				T. Arima, R. A. Drewell, K. L. Arney, J. Inoue, Y. Makita, A. Hata, M. Oshimura, N. Wake, and M. A. Surani A conserved imprinting control region at the HYMAI/ZAC domain is implicated in transient neonatal diabetes mellitus Hum. Mol. Genet., July 1, 2001; 10(14): 1475 - 1483. [Abstract] [Full Text] [PDF]

				A. C. Bell, A. G. West, and G. Felsenfeld Insulators and Boundaries: Versatile Regulatory Elements in the Eukaryotic Genome Science, January 19, 2001; 291(5503): 447 - 450. [Abstract] [Full Text]

				J. Ainscough, R. John, S. Barton, and M. Surani A skeletal muscle-specific mouse Igf2 repressor lies 40 kb downstream of the gene Development, January 9, 2000; 127(18): 3923 - 3930. [Abstract] [PDF]