Biological significancy of epigenetic modification in germ line genome

种系基因组表观遗传修饰的生物学意义

• 批准号: 16510146
• 负责人: NOZAKI Masami
• 金额: $ 2.5万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16510146/
• 关键词: Germ cell; Spermatogenesis; Genome; Epigenetics; DNA methylation; Haploid; Spermatozoa; メチル化; 遺伝子; ヒストン; プロタミン; マウス; CpGアイランド

The haploid spermatid specific gene, Tact1, is a single exon gene, which contains CpG-island within the gene. DNA methylation in the CpG-island represses the Tact1 gene expression in somatic cells. Demethylation is necessary for Tact1 expression in the round spermatids. In this study, we examined the timing, in which these CpGs are demethylated and remethylated, and involvement of the de novo methyltransferases, Dnmt3a and Dnmt3b with remethylation. We performed bisulfite-PCR sequencing for analyzing methylation status in the CpG sites within the Tact1 gene using genomic DNAs obtained from spermatogenic cells of various differentiation stages by laser micro dissection and elutriation. Demethylation occurs during proliferation stage of the spermatogonia, resulting from passive action due to inactivation of maintenance methyltransferase, Dnmt1. Hypomethylated status is maintained in pachytene spermatocytes, round spermatids, in which the Tact1 gene is expressed, and spermatozoa in epididymis. We found that remethylation occurs in mature spermatozoa in vas deferens. Analysis of germ line specific conditional knockout mice showed that genomic remethylation in spermatozoa may be redundantly regulated by two de novo methyltransferases, Dnmt3a and Dnmt3b. Alternatively, unidentified de novo methylation machinery may be necessary for newly methylation in sperm.

单倍体精子特异性基因TACT1是单个外显子基因，该基因在基因内包含CpG-Island。 CpG-Island中的DNA甲基化抑制体细胞中的TACT1基因表达。脱甲基化对于圆形精子中的TACT1表达是必需的。在这项研究中，我们检查了这些CPG被脱甲基化和二甲基化的时机，并参与了从头甲基转移酶，DNMT3A和DNMT3B，并以re甲基化。我们使用从各种分化阶段通过激光微剖析和消除率从各种分化阶段获得的基因组DNA进行了Bisulfite-PCR测序，以分析TACK1基因中CpG位点中的甲基化状态。脱甲基化发生在精子的增殖阶段，这是由于维持甲基转移酶DNMT1导致的被动作用而导致的。降压甲基化状态保持在pachytene精子细胞，圆形精子中，其中表达tACT1基因，而在附子症中的精子。我们发现，在VAS延期的成熟精子中发生了甲基化。分析种系特异性条件敲除小鼠，表明精子中的基因组再甲基化可能受到两种从头甲基转移酶DNMT3A和DNMT3B的冗余调节。或者，对于精子新近甲基化，可能需要使用未识别的从头甲基化机制。

项目成果

Genes expressing in the testis

睾丸中表达的基因

• 发表时间: 2004
• 期刊: Hormone Frontier in Gynecology 11
• 作者: Tanabe;H.;Amano;M.;Chiba;M.;Masami Nozaki
• 通讯作者: Masami Nozaki

Structure and evolution of testicular germ cell-specific genes, Oxct2a and Oxct2b

睾丸生殖细胞特异性基因 Oxct2a 和 Oxct2b 的结构和进化

• 发表时间: 2004
• 期刊: Genomics 83
• 作者: Yamada H;Sugimura H et al.;日浦 啓全;Masayoshi Onishi
• 通讯作者: Masayoshi Onishi

The adaptor molecule FADD from Xenopus laevis demonstrates evolution of its pro-apoptotic activity

来自非洲爪蟾的接头分子 FADD 展示了其促凋亡活性的进化

• 发表时间: 2004
• 期刊: Genes to Cells 9
• 作者: 田辺秀之;望月由子;千葉磨玲;宮原 正明;Kazuhiro Sakamaki
• 通讯作者: Kazuhiro Sakamaki

cAMP-responsive element in TATA-less core promoter is essential for haploid-specific gene expression in mouse testis

• DOI: 10.1093/nar/gki652
• 发表时间: 2005-01-01
• 期刊: NUCLEIC ACIDS RESEARCH
• 影响因子: 14.9
• 作者: Somboonthum, P;Ohta, H;Nozaki, M
• 通讯作者: Nozaki, M

Recent Research Developments in Biochemistry

生物化学的最新研究进展

• 发表时间: 2006
• 作者: Nozaki;M.;Hisano;M.
• 通讯作者: M.