RRC ID 32638
Author Kurihara M, Shiraishi A, Satake H, Kimura AP.
Title 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.
Journal J Mol Biol
Abstract 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 3.4kb 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.
Volume 426(17)
Pages 3069-93
Published 2014-8-26
DOI 10.1016/j.jmb.2014.06.018
PII S0022-2836(14)00334-9
PMID 25020229
MeSH Animals Base Sequence Cell Adhesion Molecules / genetics* Cell Adhesion Molecules / metabolism Chromosomal Proteins, Non-Histone / genetics Conserved Sequence Gene Expression Regulation* Histones / metabolism Liver / metabolism Male Mice Mice, Inbred C57BL Molecular Sequence Data Muscle Proteins / genetics NIH 3T3 Cells Organ Specificity Promoter Regions, Genetic Protein Processing, Post-Translational RNA, Long Noncoding / genetics* RNA, Long Noncoding / metabolism Spermatocytes / metabolism
IF 4.76
Times Cited 34
DNA material B6N BAC Mouse (RDB07573) B6Ng01-276I01
Human and Animal Cells NIH3T3-3-4(RCB1862) Hepa 1-6(RCB1638)