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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以用作特异性细胞特异性增强子和双向促进剂，用于普遍地表达基因和睾丸特异性的长度非分量RNA

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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被各种元素如促进剂，增强剂和长的非分量RNA（LNCRNA）紧密调节。在本研究中，我们将保守的非编码序列（CNS1）鉴定为精胶细胞特异性小鼠睾丸粘附分子1（TCAM1）基因的新型增强剂。 CNS1位于TCAM1基因上游的3.4kb，并与睾丸生殖细胞中的组蛋白H3K4单甲基化相关。通过体外记者基因测定，CNS1仅能够在GC-2SPD（TS）细胞中增强TCAM1启动子活性，所述GC-2SPD（TS）细胞衍生自小鼠精子细胞。当我们整合有或没有将CNS1删除与增强的绿色荧光蛋白基因的CNS1缺失到GC-2SPD（TS）细胞的染色质中，CNS1显着增强了TCAM1启动子活性的6.9-Kb 5'侧序列。这些结果表明CNS1可以用作特异性细胞特异性增强剂。有趣的是，CNS1还在报告试验中显示出高双向启动子活性，并一致，SMARCD2基因和LNCRNA指定的LNCRNA-TCAM1从CNS1的相邻区域转录。虽然SMARCD2普遍表达，但LNCRNA-TCAM1表达仅限于睾丸生殖细胞，尽管该LNCRNA未参与TCAM1活化。无处不在的SMARCD2表达与CNS1的CPG哌甲基化相关并由SP1部分控制。然而，对于LNCRNA-TCAM1转录，还似乎需要与组蛋白乙酰化和组蛋白H3K4三甲基化的强关系。本数据表明CNS1是用于TCAM1基因的精细性增强剂和SMARCD2和LNCRNA-TCAM1的双向启动子。

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