Transient homologous chromosome pairing marks the onset of X inactivation

Mammalian X inactivation turns off one female X chromosome to enact dosage compensation between XX and XY individuals. X inactivation is known to be regulated in cis by Xite, Tsix, and Xist, but in principle the two Xs must also be regulated in trans to ensure mutually exclusive silencing. Here, we demonstrate that interchromosomal pairing mediates this communication. Pairing occurs transiently at the onset of X inactivation and is specific to the X-inactivation center. Deleting Xite and Tsix perturbs pairing and counting/choice, whereas their autosomal insertion induces de novo X-autosome pairing. Ectopic X-autosome interactions inhibit endogenous X-X pairing and block the initiation of X-chromosome inactivation. Thus, Tsix and Xite function both in cis and in trans. We propose that Tsix and Xite regulate counting and mutually exclusive choice through X-X pairing.     

Molecular coupling of Xist regulation and pluripotency

During mouse embryogenesis, reversion of imprinted X chromosome inactivation in the pluripotent inner cell mass of the female blastocyst is initiated by the repression of Xist from the paternal X chromosome. Here we report that key factors supporting pluripotency-Nanog, Oct3/4, and Sox2-bind within Xist intron 1 in undifferentiated embryonic stem (ES) cells. Whereas Nanog null ES cells display a reversible and moderate up-regulation of Xist in the absence of any apparent modification of Oct3/4 and Sox2 binding, the drastic release of all three factors from Xist intron 1 triggers rapid ectopic accumulation of Xist RNA. We conclude that the three main genetic factors underlying pluripotency cooperate to repress Xist and thus couple X inactivation reprogramming to the control of pluripotency during embryogenesis.     

Epigenetic dynamics of imprinted X inactivation during early mouse development

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 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.     

非编码RNA Xist调节X染色体失活的分子机制

非编码KNAXist介导的X染色体失活是表观遗传研究的一个典范,该系统能使一整条染色体变为异染状态。从Xist与X染色体计数、失活的选择、失活的起始和失活的维持等方面综述了其分子机制。     

An X chromosome gene, WTX, is commonly inactivated in Wilms tumor

Wilms tumor is a pediatric kidney cancer associated with inactivation of the WT1 tumor-suppressor gene in 5 to 10% of cases. Using a high-resolution screen for DNA copy-number alterations in Wilms tumor, we identified somatic deletions targeting a previously uncharacterized gene on the X chromosome. This gene, which we call WTX, is inactivated in approximately one-third of Wilms tumors (15 of 51 tumors). Tumors with mutations in WTX lack WT1 mutations, and both genes share a restricted temporal and spatial expression pattern in normal renal precursors. In contrast to biallelic inactivation of autosomal tumor-suppressor genes, WTX is inactivated by a monoallelic "single-hit" event targeting the single X chromosome in tumors from males and the active X chromosome in tumors from females.     

X-Chromosome inactivation in cloned mouse embryos

To study whether cloning resets the epigenetic differences between the two X chromosomes of a somatic female nucleus, we monitored X inactivation in cloned mouse embryos. Both X chromosomes were active during cleavage of cloned embryos, followed by random X inactivation in the embryo proper. In the trophectoderm (TE), X inactivation was nonrandom with the inactivated X of the somatic donor being chosen for inactivation. When female embryonic stem cells with two active X chromosomes were used as donors, random X inactivation was seen in the TE and embryo. These results demonstrate that epigenetic marks can be removed and reestablished on either X chromosome during cloning. Our results also suggest that the epigenetic marks imposed on the X chromosomes during gametogenesis, responsible for normal imprinted X inactivation in the TE, are functionally equivalent to the marks imposed on the chromosomes during somatic X inactivation.     

Expression of the mammalian X chromosome before and after fertilization

The activity of hypoxanthine-guanine phosphoribosyltransferase in unfertilized mouse ova and in mouse embryos at the two-cell stage is proportional to the number of X chromosomes present during oogenesis. This indicates that the enzyme is X-linked in the mouse and that inactivation of the X chromosome does not occur during oogenesis. However, the genetic dosage effect of the X chromosomes is not present after the increase in hypoxanthine-guanine phosphoribosyltransferase activity in the late morula and the blastocyst stages. These results indicate that the X-linked enzyme lacuts is expressed sometimne after fertilization but before the morula stage.     

Reactivation of the paternal X chromosome in early mouse embryos

It is generally accepted that paternally imprinted X inactivation occurs exclusively in extraembryonic lineages of mouse embryos, whereas cells of the embryo proper, derived from the inner cell mass (ICM), undergo only random X inactivation. Here we show that imprinted X inactivation, in fact, occurs in all cells of early embryos and that the paternal X is then selectively reactivated in cells allocated to the ICM. This contrasts with more differentiated cell types where X inactivation is highly stable and generally irreversible. Our observations illustrate that an important component of genome plasticity in early development is the capacity to reverse heritable gene silencing decisions.     

Genetics. Reprogramming X inactivation

Inactivation of one of the two X chromosomes occurs in all cells of female adult mice so that genes are expressed from only one X chromosome. In a Perspective, Clerc and Avner describe an elegant series of experiments in mouse embryos cloned from adult and embryonic female cell nuclei (Eggan et al.) that reveal how the inactivation state of the X chromosomes is reprogrammed.     

Mammalian X-chromosome action: inactivation limited in spread and region of origin

In its simplest form the hypothesis of the single-active-X chromosome does not explain variegated-type position effects in the mouse. Inactivity appears not to involve one entire X chromosome; furthermore, even those parts of the chromosome that can change to an inactive state spread inactivation not to the entire attached piece of autosome, but along a gradient to limited distances.     

A chicken transferrin gene in transgenic mice escapes X-chromosome inactivation

Mammalian X-chromosome inactivation involves a coordinate shutting down of physically linked genes. Several proposed models require the presence of specific sequences near genes to permit the spread of inactivation into these regions. If such models are correct, one might predict that heterologous genes transferred onto the X chromosome might lack the appropriate signal sequences and therefore escape inactivation. To determine whether a foreign gene inserted into the X chromosome is subject to inactivation, transgenic mice harboring 11 copies of the complete, 17-kilobase chicken transferrin gene on the X chromosome were used. Male mice hemizygous for this insert were bred with females bearing Searle's translocation, an X-chromosome rearrangement that is always active in heterozygous females (the unrearranged X chromosome is inactive). Female offspring bearing the Searle's translocation and the chicken transferrin gene had the same amount of chicken transferrin messenger RNA in liver as did transgenic male mice or transgenic female mice lacking the Searle's chromosome. This result shows that the inserted gene is not subject to X-chromosome inactivation and suggests that the inactivation process cannot spread over 187 kilobases of DNA in the absence of specific signal sequences required for inactivation.     

Senescence of nickel-transformed cells by an X chromosome: possible epigenetic control

Transfer of a normal Chinese hamster X chromosome (carried in a mouse A9 donor cell line) to a nickel-transformed Chinese hamster cell line with an Xq chromosome deletion resulted in senescense of these previously immortal cells. At early passages of the A9/CX donor cells, the hamster X chromosome was highly active, inducing senescence in 100% of the colonies obtained after its transfer into the nickel-transformed cells. However, senescence was reduced to 50% when Chinese hamster X chromosomes were transferred from later passage A9 cells. Full senescing activity of the intact hamster X chromosome was restored by treatment of the donor mouse cells with 5-azacytidine, which induced demethylation of DNA. These results suggest that a senescence gene or genes, which may be located on the Chinese hamster X chromosome, can be regulated by DNA methylation, and that escape from senescence and possibly loss of tumor suppressor gene activity can occur by epigenetic mechanisms.     

Mammalian oocytes: X chromosome activity

The glucose-6-phosphate dehydrogenase and lactate dehydrogenase contents of oocytes from XO and XX female mice have been measured. The activity of the former in the oocytes of XO mice is half of that in the oocytes of XX mice, whereas the lactate dehydrogenase activities in the two groups of ova are the same. These results indicate that glucose-6-phosphate dehydrogenase from a mouse oocyte is an X-linked enzyme, that its synthesis occurs in the oocyte and is dosage dependent, and that inactivation of the X chromosome does not occur in the mouse oocyte.     

Human phosphoglycerate kinase and inactivation of the X chromosome

The fibroblasts derived from the skin of a woman heterozygous for an X-linked deficiency of phosphoglycerate kinase represented a mosaic. Two of 22 clones with normal glucose-6-phosphate dehydrogenase activity and hypoxanthine(guanine) phosphoribosyltransferase activity had no phosphoglycerate kinase activity detected by electrophoresis. Because the loci for glucose-6-phosphate dehydrogeniase and hypoxanthine(guanine)phosphoribosyltransferase are already known to undergo inactivation and to be on the short arm of the X chromosome and the locus for phosphoglycerate kinase is on the long arm, these observations support the conclusion that the entire human X chromosome can be involved in X inactivation.     

Role of histone H3 lysine 27 methylation in X inactivation

The Polycomb group (PcG) protein Eed is implicated in regulation of imprinted X-chromosome inactivation in extraembryonic cells but not of random X inactivation in embryonic cells. The Drosophila homolog of the Eed-Ezh2 PcG protein complex achieves gene silencing through methylation of histone H3 on lysine 27 (H3-K27), which suggests a role for H3-K27 methylation in imprinted X inactivation. Here we demonstrate that transient recruitment of the Eed-Ezh2 complex to the inactive X chromosome (Xi) occurs during initiation of X inactivation in both extraembryonic and embryonic cells and is accompanied by H3-K27 methylation. Recruitment of the complex and methylation on the Xi depend on Xist RNA but are independent of its silencing function. Together, our results suggest a role for Eed-Ezh2-mediated H3-K27 methylation during initiation of both imprinted and random X inactivation and demonstrate that H3-K27 methylation is not sufficient for silencing of the Xi.     

Extensive gene traffic on the mammalian X chromosome

Mammalian sex chromosomes have undergone profound changes since evolving from ancestral autosomes. By examining retroposed genes in the human and mouse genomes, we demonstrate that, during evolution, the mammalian X chromosome has generated and recruited a disproportionately high number of functional retroposed genes, whereas the autosomes experienced lower gene turnover. Most autosomal copies originating from X-linked genes exhibited testis-biased expression. Such export is incompatible with mutational bias and is likely driven by natural selection to attain male germline function. However, the excess recruitment is consistent with a combination of both natural selection and mutational bias.     

Mosaic histocompatibility of skin grafts from female mice

The histoincompatibility determined by one or more genes on the X chromosome of the mouse effects a complete rejection of skin of the (C57BL/6 female symbol x BALB/c male symbol) F(1) hybrid male grafted onto the reciprocal type F(1) hybrid male, but only an incomplete rejection of either reciprocal type F(1) hybrid female skin, grafted onto the same type of male host. The resulting mosaic survival pattern of the female graft is interpreted as support for the Lyon hypothesis of X-chromosome inactivation.     

Chromosomal localization of muscle nicotinic acetylcholine receptor genes in the mouse

The chromosomal localization of the genes encoding the four subunits of muscle nicotinic receptor was determined by analyzing restriction fragment length polymorphisms between two mouse species Mus musculus domesticus (DBA/2) and Mus spretus (SPE). Analysis of the progeny of the interspecies mouse backcross (DBA/2 X SPE) X DBA/2 showed that the alpha-subunit gene cosegregates with the alpha-cardiac actin gene on chromosome 17, that the beta-subunit gene is located on chromosome 11, and that the gamma- and delta-subunit genes cosegregate and are located on chromosome 1.     

海门白山羊染色体核型研究及C-带分析

采用外周血淋巴细胞培养及染色体分带技术,分析了海门白山羊的染色体核型与C-带。结果表明,海门白山羊二倍体染色体数为2n=60,常染色体和X染色体均为端部着丝粒染色体,X染色体的大小介于1号和2号染色体之间,Y染色体最小,为中部着丝粒染色体,公羊核型为60,XY,母羊为60,XX。大部分常染色体和X,Y染色体着丝粒部位显示阳性C-带,但不同染色体的阳性C-带区域大小不同。