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首页> 外文期刊>Molecular and Cellular Biology >A Functional Chromatin Domain Does Not Resist X Chromosome Inactivation: Silencing of cLys Correlates with Methylation of a Dual Promoter-Replication Origin
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A Functional Chromatin Domain Does Not Resist X Chromosome Inactivation: Silencing of cLys Correlates with Methylation of a Dual Promoter-Replication Origin

机译：功能性染色质域不能抵抗X染色体失活：cLys沉默与双启动子复制起源的甲基化相关。

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摘要

To investigate the molecular mechanism(s) involved in the propagation and maintenance of X chromosome inactivation (XCI), the 21.4-kb chicken lysozyme (cLys) chromatin domain was inserted into the Hprt locus on the mouse X chromosome. The inserted fragment includes flanking matrix attachment regions (MARs), an origin of bidirectional replication (OBR), and all the cis-regulatory elements required for correct tissue-specific expression of cLys. It also contains a recently identified and widely expressed second gene, cGas41. The cLys domain is known to function as an autonomous unit resistant to chromosomal position effects, as evidenced by numerous transgenic mouse lines showing copy-number-dependent and development-specific expression of cLys in the myeloid lineage. We asked the questions whether this functional chromatin domain was resistant to XCI and whether the X inactivation signal could spread across an extended region of avian DNA. A generally useful method was devised to generate pure populations of macrophages with the transgene either on the active (Xa) or the inactive (Xi) chromosome. We found that (i) cLys and cGas41 are expressed normally from the Xa; (ii) the cLys chromatin domain, even when bracketed by MARs, is not resistant to XCI; (iii) transcription factors are excluded from lysozyme enhancers on the Xi; and (iv) inactivation correlates with methylation of a CpG island that is both an OBR and a promoter of the cGas41 gene.
机译：为了研究X染色体失活（XCI）的传播和维持的分子机制，将21.4kb鸡溶菌酶（ Lys ）染色质结构域插入到 Hprt 小鼠X染色体上的基因座。插入的片段包括侧翼基质附着区（MARs），双向复制（OBR）的起源以及正确表达组织特异性表达 cLys cis 调控元件>。它还包含最近鉴定并广泛表达的第二个基因 cGas41 。已知 cLys 结构域可作为抵抗染色体位置效应的自主单位，许多转基因小鼠品系均显示了 cLys 在骨髓谱系中。我们询问了以下问题：该功能性染色质结构域是否对XCI具有抗性，X灭活信号是否可以在禽类DNA的扩展区域中传播。设计了一种通常有用的方法，以在活动（Xa）或非活动（Xi）染色体上生成带有转基因的纯巨噬细胞群体。我们发现（i） cLys 和 cGas41 是从Xa正常表达的; （ii）即使在MARs包围下， cLys 染色质结构域也不抗XCI; （iii）从Xi上的溶菌酶增强子中排除转录因子; （iv）失活与CpG岛的甲基化有关，该岛既是OBR，又是 cGas41 基因的启动子。

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