Abstract

Oog1 is an oocyte-specific gene whose expression is turned on in mouse oocytes at embryonic day (E) 15.5, concomitant with the time when most of the female germ cells stop proliferating and enter meiotic prophase. Here, we characterize the Oog1 promoter, and show that transgenic GFP reporter expression driven by the 2.7 kb and 3.9 kb regions upstream of the Oog1 transcription start site recapitulates the intrinsic Oog1 expression pattern. In addition, the 3.9 kb upstream region exhibits stronger transcriptional activity than does the 2.7 kb region, suggesting that regulatory functions might be conserved in the additional 1.2 kb region found within the 3.9 kb promoter. Interestingly, the longer promoter (3.9 kb) also showed strong activity in male germ cells, from late pachytene spermatocytes to elongated spermatids. This is likely due to the aberrant demethylation of two CpG sites in the proximal promoter region. One was highly methylated in the tissues in which GFP expression was suppressed, and another was completely demethylated only in Oog1pro3.9 male and female germ cells. These results suggest that aberrant demethylation of the proximal promoter region induced ectopic expression in male germ cells under the control of 3.9 kb Oog1 promoter. This is the first report indicating that sex-dependent gene expression is altered according to the length and the methylation status of the promoter region. Additionally, our results show that individual CpG sites are differentially methylated and play different roles in regulating promoter activity and gene transcription.

Highlights

  • Oogenesin1 (Oog1) is an oocyte-specific gene that is expressed after entry into meiosis and during early embryogenesis [1]

  • We found that the 3.9 kb promoter functioned in male germ cells, and that the methylation status of the proximal promoter region differed between the Oog1pro2.7 and Oog1pro3.9 transgenes in male and female germ cells, suggesting that CpG methylation of the proximal region of the Oog1 promoter may control gene expression in both male and female germ cells

  • We compared the upstream regions of all five copies of Oog1 to identify the promoter region

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Summary

Introduction

Oogenesin (Oog1) is an oocyte-specific gene that is expressed after entry into meiosis and during early embryogenesis [1]. The mouse genome contains five copies of Oog clustered on chromosomes 4 and 12. All of the copies contain a TATA-box in the proximal upstream region, suggesting that they are transcribed. Oog expression begins in oocytes at E15.5 and continues to the 2-cell stage following fertilization [1]. OOG1 protein is localized in the nucleus of late 1-cell to early 2-cell embryos, concomitant with zygotic gene activation and first mitotic division. We previously identified a potential binding partner of OOG1, Ras and Ral guanine nucleotide dissociation stimulator (RalGDS), by yeast two-hybrid screening of a germinal vesicle (GV) oocyte cDNA library [2], but the function of Oog remains unknown

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