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首页> 外文期刊>Journal of Molecular Biology >A conserved noncoding sequence can function as a spermatocyte-specific enhancer and a bidirectional promoter for a ubiquitously expressed gene and a testis-specific long noncoding RNA
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A conserved noncoding sequence can function as a spermatocyte-specific enhancer and a bidirectional promoter for a ubiquitously expressed gene and a testis-specific long noncoding RNA

机译：保守的非编码序列可作为精子细胞特异性增强子和遍在表达基因和睾丸特异性长非编码RNA的双向启动子

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摘要

Tissue-specific gene expression is tightly regulated by various elements such as promoters, enhancers, and long noncoding RNAs (lncRNAs). In the present study, we identified a conserved noncoding sequence (CNS1) as a novel enhancer for the spermatocyte-specific mouse testicular cell adhesion molecule 1 (Tcam1) gene. CNS1 was located 3.4 kb upstream of the Tcam1 gene and associated with histone H3K4 mono-methylation in testicular germ cells. By the in vitro reporter gene assay, CNS1 could enhance Tcam1 promoter activity only in GC-2spd(ts) cells, which were derived from mouse spermatocytes. When we integrated the 6.9-kb 5′-flanking sequence of Tcam1 with or without a deletion of CNS1 linked to the enhanced green fluorescent protein gene into the chromatin of GC-2spd(ts) cells, CNS1 significantly enhanced Tcam1 promoter activity. These results indicate that CNS1 could function as a spermatocyte-specific enhancer. Interestingly, CNS1 also showed high bidirectional promoter activity in the reporter assay, and consistent with this, the Smarcd2 gene and lncRNA, designated lncRNA-Tcam1, were transcribed from adjacent regions of CNS1. While Smarcd2 was ubiquitously expressed, lncRNA-Tcam1 expression was restricted to testicular germ cells, although this lncRNA did not participate in Tcam1 activation. Ubiquitous Smarcd2 expression was correlated to CpG hypo-methylation of CNS1 and partially controlled by Sp1. However, for lncRNA-Tcam1 transcription, the strong association with histone acetylation and histone H3K4 tri-methylation also appeared to be required. The present data suggest that CNS1 is a spermatocyte-specific enhancer for the Tcam1 gene and a bidirectional promoter of Smarcd2 and lncRNA-Tcam1.
机译：组织特异性基因表达受到各种元素的严格调控，例如启动子，增强子和长非编码RNA（lncRNA）。在本研究中，我们确定了保守的非编码序列（CNS1）作为精子细胞特异性小鼠睾丸细胞粘附分子1（Tcam1）基因的新型增强子。 CNS1位于Tcam1基因上游3.4 kb，并与睾丸生殖细胞中的组蛋白H3K4单甲基化有关。通过体外报告基因测定，CNS1仅在源自小鼠精母细胞的GC-2spd（ts）细胞中才能增强Tcam1启动子活性。当我们将Tcam1的6.9-kb 5'侧翼序列整合有缺失或缺失与增强型绿色荧光蛋白基因相连的CNS1到GC-2spd（ts）细胞的染色质中时，CNS1显着增强了Tcam1启动子活性。这些结果表明CNS1可以作为精母细胞特异性增强子。有趣的是，CNS1在报告基因检测中也显示出较高的双向启动子活性，与此相符的是，从CNS1的相邻区域转录了Smarcd2基因和lncRNA，称为lncRNA-Tcam1。虽然Smarcd2被普遍表达，但lncRNA-Tcam1的表达仅限于睾丸生殖细胞，尽管该lncRNA不参与Tcam1的激活。普遍存在的Smarcd2表达与CNS1的CpG低甲基化相关，并部分受Sp1控制。然而，对于lncRNA-Tcam1转录，似乎也需要与组蛋白乙酰化和组蛋白H3K4三甲基化的强烈关联。本数据表明CNS1是Tcam1基因的精母细胞特异性增强子，是Smarcd2和lncRNA-Tcam1的双向启动子。

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