Team Publications Private: Mammalian Developmental Epigenetics

Year of publication 2006

Christian P Bacher, Michèle Guggiari, Benedikt Brors, Sandrine Augui, Philippe Clerc, Philip Avner, Roland Eils, Edith Heard (2006 Jan 26) Transient colocalization of X-inactivation centres accompanies the initiation of X inactivation. Nature cell biology : 293-9

Summary

The initial differential treatment of the two X chromosomes during X-chromosome inactivation is controlled by the X-inactivation centre (Xic). This locus determines how many X chromosomes are present in a cell (‘counting’) and which X chromosome will be inactivated in female cells (‘choice’). Critical control sequences in the Xic include the non- coding RNAs Xist and Tsix, and long-range chromatin elements. However, little is known about the process that ensures that X inactivation is triggered appropriately when more than one Xic is present in a cell. Using three-dimensional fluorescence in situ hybridization (FISH) analysis, we showed that the two Xics transiently colocalize, just before X inactivation, in differentiating female embryonic stem cells. Using Xic transgenes capable of imprinted but not random X inactivation, and Xic deletions that disrupt random X inactivation, we demonstrated that Xic colocalization is linked to Xic function in random X inactivation. Both long-range sequences and the Tsix element, which generates the antisense transcript to Xist, are required for the transient interaction of Xics. We propose that transient colocalization of Xics may be necessary for a cell to determine Xic number and to ensure the correct initiation of X inactivation.

Year of publication 2005

Ikuhiro Okamoto, Danielle Arnaud, Patricia Le Baccon, Arie P Otte, Christine M Disteche, Philip Avner, Edith Heard (2005 Oct 18) Evidence for de novo imprinted X-chromosome inactivation independent of meiotic inactivation in mice. Nature : 369-73

Summary

In mammals, one of the two X chromosomes is inactivated in females to enable dosage compensation for X-linked gene products. In rodents and marsupials, only the X chromosome of paternal origin (Xp) is silenced during early embryogenesis. This could be due to a carry- over effect of the X chromosome’s passage through the male germ line, where it becomes transiently silenced together with the Y chromosome, during meiotic sex chromosome inactivation (MSCI). Here we show that Xist (X inactive specific transcript) transgenes, located on autosomes, do not undergo MSCI in the male germ line of mice and yet can induce imprinted cis-inactivation when paternally inherited, with identical kinetics to the Xp chromosome. This suggests that MSCI is not necessary for imprinted X-chromosome

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inactivation in mice. We also show that the Xp is transcribed, like autosomes, at zygotic gene activation rather than being ‘pre-inactivated’. We propose that expression of the paternal Xist gene at zygotic gene activation is sufficient to trigger cis-inactivation of the X chromosome, or of an autosome carrying a Xist transgene.

Edith Heard (2005 Aug 19) Delving into the diversity of facultative heterochromatin: the epigenetics of the inactive X chromosome. Current opinion in genetics & development : 482-9

Summary

X chromosome inactivation represents one of the most dramatic examples of mono-allelic gene expression and long-term gene-silencing in mammals. The key regulatory molecule that triggers silencing is the Xist transcript, but little is known about its repressive action. Some progress has been made in deciphering the epigenetics of the inactive state that it triggers, however. During pre-implantation development, the inactive state is relatively labile. Later on, in the soma, the inactive state is highly stable and clonally heritable. This is ensured by the panoply of epigenetic modifications that characterize the inactive X and, presumably, is also a result of its spatio-temporal segregation. The inactive X chromosome has been associated with an increasing number of histone modifications, and several recent studies have implicated Polycomb group proteins in laying down some of these marks. Thanks to genetic and biochemical approaches to analyse these proteins, the epigenetic tapestry of the inactive X is just beginning to be unravelled. Lineage-specific differences provide a glimpse into the developmental complexity of the epigenetic marks that ensure the inactive state.

Year of publication 2004

Claire Rougeulle, Julie Chaumeil, Kavitha Sarma, C David Allis, Danny Reinberg, Philip Avner, Edith Heard (2004 Jun 1) Differential histone H3 Lys-9 and Lys-27 methylation profiles on the X chromosome. Molecular and cellular biology : 5475-84

Summary

Histone H3 tail modifications are among the earliest chromatin changes in the X- chromosome inactivation process. In this study we investigated the relative profiles of two important repressive marks on the X chromosome: methylation of H3 lysine 9 (K9) and 27 (K27). We found that both H3K9 dimethylation and K27 trimethylation characterize the inactive X in somatic cells and that their relative kinetics of enrichment on the X chromosome as it undergoes inactivation are similar. However, dynamic changes of H3K9 and H3K27 methylation on the inactivating X chromosome compared to the rest of the

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genome are distinct, suggesting that these two modifications play complementary and perhaps nonredundant roles in the establishment and/or maintenance of X inactivation. Furthermore, we show that a hotspot of H3K9 dimethylation 5′ to Xist also displays high levels of H3 tri-meK27. However, analysis of this region in G9a mutant embryonic stem cells shows that these two methyl marks are dependent on different histone methyltransferases.

Edith Heard (2004 May 18) Recent advances in X-chromosome inactivation. Current opinion in cell biology : 247-55

Summary

X inactivation is the silencing one of the two X chromosomes in XX female mammals. Initiation of this process during early development is controlled by the X-inactivation centre, a complex locus that determines how many, and which, X chromosomes will be inactivated. It also produces the Xist transcript, a remarkable RNA that coats the X chromosome in cis and triggers its silencing. Xist RNA coating induces a cascade of chromatin changes on the X chromosome, including the recruitment of Polycomb group proteins. This results in an inactive state that is initially labile, but may be further locked in by epigenetic marks such as DNA methylation. In mice, X inactivation has recently been found to be much more dynamic than previously thought during early pre-implantation development. The paternal X chromosome is initially inactivated in all cells of cleavage-stage embryos and then selectively reactivated in the subset of cells that will form the embryo, with random X inactivation occurring thereafter.

Julie Chaumeil, Ikuhiro Okamoto, Edith Heard (2004 Feb 21) X-chromosome inactivation in mouse embryonic stem cells: analysis of histone modifications and transcriptional activity using immunofluorescence and FISH. Methods in enzymology : 405-19

Summary

Year of publication 2003

Ikuhiro Okamoto, Arie P Otte, C David Allis, Danny Reinberg, Edith Heard (2003 Dec 13) Epigenetic dynamics of imprinted X inactivation during early mouse development. Science (New York, N.Y.) : 644-9

Summary

The initiation of X-chromosome inactivation is thought to be tightly correlated with early differentiation events during mouse development. Here, we show that although initially

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active, the paternal X chromosome undergoes imprinted inactivation from the cleavage stages, well before cellular differentiation. A reversal of the inactive state, with a loss of epigenetic marks such as histone modifications and polycomb proteins, subsequently occurs in cells of the inner cell mass (ICM), which give rise to the embryo-proper in which random X inactivation is known to occur. This reveals the remarkable plasticity of the X-inactivation process during preimplantation development and underlines the importance of the ICM in global reprogramming of epigenetic marks in the early embryo.

Year of publication 2002

Claire Rougeulle, Edith Heard (2002 Aug 15) Antisense RNA in imprinting: spreading silence through Air. Trends in genetics : TIG : 434-7

Summary

In some animals, including mammals, a number of genes are expressed differently according to whether they have been inherited from the mother or from the father, through a process known as genomic imprinting. Noncoding RNAs have increasingly been found associated with imprinted genes, but their role, if any, has remained enigmatic. A recent study provides the first evidence that, at least in one case, a noncoding RNA has a direct role in regulating imprinted gene expression in cis.

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