Molecular mechanism of spermatogenesis regulated by a testis-specific poly(A) polymerase, TPAP

睾丸特异性多聚腺苷酸聚合酶TPAP调节精子发生的分子机制

• 批准号: 15380068
• 负责人: KASHIWABARA Shin-ichi
• 金额: $ 8.06万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380068/
• 关键词: Spermatogenesis; Poly(A) chain; RNA-binding protein; Protein trafficking; TPAP; Cytoplasmic polyadenylation; mRNA; 細胞質ポリアデニル化反応; モータータンパク質; トランスジェニックマウス; KIF2A

To elucidate the molecular mechanism of spermatogenesis, I have examined the implication of a testis-specific poly(A) polymerase, TPAP, in mouse spermatogenesis. The following experimental results have been obtained :1.Transgenic mice overexpressing TPAP exhibited normal spermatogenesis and fertility. The sizes of some transcription factor mRNAs as the substrates of TPAP were also unaffected, implying the presence of a regulatory mechanism(s) defining the extent of the cytoplasmic mRNA polyadenylation.2.TPAP was capable of interacting with mRNAs encoding germ cell morphogenesis-regulating proteins and transcription factors, including protamine, TFIIAγ, and CREMτ. These data suggest that TPAP presumably has the selectivity for mRNA recognition.3.Cytoplasmic polyadenylation element-binding protein 2, CPEB2, was expressed in the testis and was capable of binding to the 3'-untranslated regions of TFIIAγ and TRF2 mRNAs, thus implying a possible implication of CPEB2 in the poly(A) elongation of mRNAs, in cooperation with TPAP.4.Poly(A) binding protein PABPC1 was capable of binding to the 3'-untranslated region of TFIIAγ mRNA only when the mRNA contained poly(A) tail. Since TPAP showed the inability to bind to the same mRNA with and without the poly(A) tail, TPAP may interact with the TFIIAγ mRNA possibly through a protein complex.5.RecQL exhibiting a helicase activity has been identified as one of the TPAP-interacting proteins. Indeed, TPAP showed a direct binding activity to RecQL.

为了阐明精子发生的分子机制，我研究了睾丸特异性聚腺苷酸聚合酶（TPAP）在小鼠精子发生中的作用。获得了以下实验结果：1.过表达TPAP的转基因小鼠具有正常的精子发生和生育能力。作为TPAP底物的一些转录因子mRNA的大小也不受影响，这意味着细胞质mRNA多聚腺苷酸化程度的调节机制的存在。2. TPAP能够与编码生殖细胞形态发生调节蛋白和转录因子的mRNA相互作用，包括鱼精蛋白、TFIIAγ和CREMτ。细胞质多聚腺苷酸化元件结合蛋白2（CPEB 2）在睾丸中表达，并能与TFIIAγ和TRF 2 mRNA的3 '端非翻译区结合，提示CPEB 2可能参与mRNA的poly（A）延伸。4. Poly（A）结合蛋白PABPC 1仅在TFIIAγ mRNA含有poly（A）尾的情况下才能与TFIIAγ mRNA的3 '非翻译区结合。由于TPAP不能与有poly（A）尾和无poly（A）尾的相同mRNA结合，TPAP可能通过蛋白复合物与TFIIAγ mRNA相互作用。5.具有解旋酶活性的RecQL被鉴定为TPAP相互作用蛋白之一。事实上，TPAP显示出与RecQL的直接结合活性。

项目成果

Transgenic expression of testis-specific poly(A) polymerase TPAP in wild-type and TPAP-deficient mice

• DOI: 10.1262/jrd.50.207
• 发表时间: 2004-04-01
• 期刊: JOURNAL OF REPRODUCTION AND DEVELOPMENT
• 影响因子: 1.8
• 作者: Zhuang, TG;Kashiwabara, S;Baba, T
• 通讯作者: Baba, T

Mouse sperm lacking ADAM1b/ADAM2 fertilin can fuse with the egg plasma membrane and effect fertilization

• DOI: 10.1074/jbc.m510558200
• 发表时间: 2006-03-03
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kim, E;Yamashita, M;Baba, T
• 通讯作者: Baba, T

Synthesis, processing, and subcellular localization of mouse ADAM3 duringa spermatogenesis and epididymal sperm transport

小鼠 ADAM3 在精子发生和附睾精子运输过程中的合成、加工和亚细胞定位

• 发表时间: 2004
• 期刊: J.Reprod.Dev. 50・5
• 作者: Kim;E.;Nishimura;H.;Iwase;S.;Yamagata;K.;Kashiwabara;S.;Baba;T.
• 通讯作者: T.

Zhuang, T., Kashiwabara, S., Noguchi, J., Baba, T.: "Transgenic expression of testis-specific poly(A) polymerase TPAP in wild-type and TPAP-deficient mice"Journal of Reproduction and Development. (in press). (2004)

Zhuang, T.、Kashiwabara, S.、Noguchi, J.、Baba, T.：“野生型和 TPAP 缺陷小鼠中睾丸特异性多聚腺苷酸聚合酶 TPAP 的转基因表达”生殖与发育杂志。

Synthesis, processing, and subcellular localization of mouse ADAM3 during spermatogenesis and epididymal sperm transport

• DOI: 10.1262/jrd.50.571
• 发表时间: 2004-10-01
• 期刊: JOURNAL OF REPRODUCTION AND DEVELOPMENT
• 影响因子: 1.8
• 作者: Kim, E;Nishimura, H;Baba, T
• 通讯作者: Baba, T