CpG site methylation regulates mouse Rec8 gene promoter activity

The Rec8 gene is specifically expressed in fetal and adult gonads. Although the importance of REC8 in gametogenesis is widely acknowledged, the mechanisms underlying its germ cell-specific expression remain unclear. In this study, we utilized the mouse Rec8 gene sequence to construct a 2577 bp seque...

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Main Authors: Mei RONG, Na FENG, Jinghuan LI, Wuyun DALAI
Format: Article
Language:English
Published: The Society for Reproduction and Development 2025-04-01
Series:The Journal of Reproduction and Development
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Online Access:https://www.jstage.jst.go.jp/article/jrd/71/3/71_2024-077/_pdf/-char/en
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author Mei RONG
Na FENG
Jinghuan LI
Wuyun DALAI
author_facet Mei RONG
Na FENG
Jinghuan LI
Wuyun DALAI
author_sort Mei RONG
collection DOAJ
description The Rec8 gene is specifically expressed in fetal and adult gonads. Although the importance of REC8 in gametogenesis is widely acknowledged, the mechanisms underlying its germ cell-specific expression remain unclear. In this study, we utilized the mouse Rec8 gene sequence to construct a 2577 bp sequence, which included intron 1 (180 bp), exon 1 (118 bp), and an upstream 2279 bp region. The dual-luciferase assay results showed significant differences in promoter activity between –650 bp and –385 bp and between –89 bp and –35 bp. This indicated that the core promoter region of the Rec8 gene may exist within these regions. Bisulfite sequencing PCR results showed that CpGs 10–19 were largely unmethylated in the testes but hypermethylated in other tissues. Interestingly, correlation analysis between CpG methylation status and Rec8 mRNA expression levels showed that methylation of CpGs 10 to 19 was negatively correlated with Rec8 mRNA expression levels (Pearson’s r = −0.991, P = 0.009). Furthermore, RNA-Seq data and bioinformatic analyses suggested that the specific expression of Rec8 may be linked to the presence of TATA-like sequences within its core promoter region. Overall, these findings indicate that Rec8 expression is regulated by the low methylation of CpG sites and the presence of TATA-like sequences in its core promoter.
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publishDate 2025-04-01
publisher The Society for Reproduction and Development
record_format Article
series The Journal of Reproduction and Development
spelling doaj-art-625a08bb4593437599bdfade3e0e68c62025-08-20T02:03:18ZengThe Society for Reproduction and DevelopmentThe Journal of Reproduction and Development0916-88181348-44002025-04-0171314515310.1262/jrd.2024-077jrdCpG site methylation regulates mouse Rec8 gene promoter activityMei RONG0Na FENG1Jinghuan LI2Wuyun DALAI3College of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, Inner Mongolia, ChinaYonghe County culture and tourism Bureau, Yonghe County, Linfen City 041400, ChinaCollege of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, Inner Mongolia, ChinaCollege of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, Inner Mongolia, ChinaThe Rec8 gene is specifically expressed in fetal and adult gonads. Although the importance of REC8 in gametogenesis is widely acknowledged, the mechanisms underlying its germ cell-specific expression remain unclear. In this study, we utilized the mouse Rec8 gene sequence to construct a 2577 bp sequence, which included intron 1 (180 bp), exon 1 (118 bp), and an upstream 2279 bp region. The dual-luciferase assay results showed significant differences in promoter activity between –650 bp and –385 bp and between –89 bp and –35 bp. This indicated that the core promoter region of the Rec8 gene may exist within these regions. Bisulfite sequencing PCR results showed that CpGs 10–19 were largely unmethylated in the testes but hypermethylated in other tissues. Interestingly, correlation analysis between CpG methylation status and Rec8 mRNA expression levels showed that methylation of CpGs 10 to 19 was negatively correlated with Rec8 mRNA expression levels (Pearson’s r = −0.991, P = 0.009). Furthermore, RNA-Seq data and bioinformatic analyses suggested that the specific expression of Rec8 may be linked to the presence of TATA-like sequences within its core promoter region. Overall, these findings indicate that Rec8 expression is regulated by the low methylation of CpG sites and the presence of TATA-like sequences in its core promoter.https://www.jstage.jst.go.jp/article/jrd/71/3/71_2024-077/_pdf/-char/encohesindna methylationgene expressionpromoterrec8
spellingShingle Mei RONG
Na FENG
Jinghuan LI
Wuyun DALAI
CpG site methylation regulates mouse Rec8 gene promoter activity
The Journal of Reproduction and Development
cohesin
dna methylation
gene expression
promoter
rec8
title CpG site methylation regulates mouse Rec8 gene promoter activity
title_full CpG site methylation regulates mouse Rec8 gene promoter activity
title_fullStr CpG site methylation regulates mouse Rec8 gene promoter activity
title_full_unstemmed CpG site methylation regulates mouse Rec8 gene promoter activity
title_short CpG site methylation regulates mouse Rec8 gene promoter activity
title_sort cpg site methylation regulates mouse rec8 gene promoter activity
topic cohesin
dna methylation
gene expression
promoter
rec8
url https://www.jstage.jst.go.jp/article/jrd/71/3/71_2024-077/_pdf/-char/en
work_keys_str_mv AT meirong cpgsitemethylationregulatesmouserec8genepromoteractivity
AT nafeng cpgsitemethylationregulatesmouserec8genepromoteractivity
AT jinghuanli cpgsitemethylationregulatesmouserec8genepromoteractivity
AT wuyundalai cpgsitemethylationregulatesmouserec8genepromoteractivity