The yeast gene controlling over the initiation of sexual reproduction

控制有性生殖起始的酵母基因

• 批准号: 03640573
• 负责人: SHIMODA Chikashi
• 金额: $ 1.54万
• 依托单位: OSAKA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03640573/
• 关键词: Fission yeast; meiosis; sexual reproduction; RNA helicase; mating; specific antibody; DEAD protein; Drosophila; ポリクローナル抗体; ウエスタンブロット; 細胞周期; 転写後調節; PCR法; 部位特異的変異誘導; RNAヘリカ-ゼ

The fission yeast ste13^+ gene which encodes a putative ATP-dependent RNA helicase is essential for initiation of sexual reproduction in response to nitrogen-starvation signal. The ste13^+ gene and its products were analyzed to verify our hypothesis that Ste13 regulates gene expression during sexual cycle at a post-transcriptional level and the following results were obtained.(1)In vitro mutagenesis of ste13 :The ste13 null mutant was defective in mating and meiosis but able to grow in rich medium. Site-directed mutagenesis introduced into the consensus motifs of RNA helicases caused the sterile phenotype. These facts indicate that the ste13 gene is not essential for mitotic cell division but indispensable for induction of sexual cycle.(2)Intracellular localization of the ste13 protein :The GST-ste13 fusion proteins were overproduced in E.coli. The affinity-purified fusion protein was used as the antigen to prepare a polyclonal antibody. Western blot analysis revealed that this antibody recognized a 58 kDa polypeptide in S.pombe cell extracts. Immunofluorescence microscopy with anti-ste13 showed that the ste13 protein was not localized in the nucleus and distributed in the cytoplasm.

裂殖酵母的ste 13 ^+基因编码一种ATP依赖的RNA解旋酶，它是响应氮饥饿信号启动有性生殖所必需的。我们对ste 13 ^+基因及其产物进行了分析，以验证我们的假设，即Ste 13在转录后水平调节性周期中的基因表达，并获得了以下结果。(1)In ste 13的体外突变：ste 13无效突变体在交配和减数分裂方面有缺陷，但能够在丰富培养基中生长。定点突变引入RNA解旋酶的共有基序引起不育表型。这些事实表明，ste 13基因是不是必不可少的有丝分裂细胞分裂，但必不可少的诱导性周期。（2）ste 13蛋白的胞内定位：GST-ste 13融合蛋白在大肠杆菌中过量表达。以亲和纯化的融合蛋白为抗原制备多克隆抗体。Western印迹分析显示，该抗体识别粟酒裂殖酵母细胞提取物中的58 kDa多肽。抗ste 13免疫荧光显微镜显示ste 13蛋白不定位于细胞核而分布于细胞质。

项目成果

T.Aono: "Mating pheromone-induced expression of the mat1-Pm gene of Schizosaccharomomyces pombe:Identification of signalling componets and characterization of upstream controlling elements" YEAST. (in press).

T.Aono：“交配信息素诱导的粟酒裂殖酵母 mat1-Pm 基因的表达：信号成分的鉴定和上游控制元件的表征”酵母。

Maekawa, H.: "The ste13^+ gene encoding a putative RNA helicase is essential for nitrogen starvation-induced G1 arrest and initiation of sexual development in the fission yeast." Mo1.Gen.Genet. (in press).

Maekawa, H.：“编码假定的 RNA 解旋酶的 ste13^ 基因对于氮饥饿诱导的裂殖酵母 G1 停滞和有性发育的启动至关重要。”

Kitamura,K.and C.Shimoda: "The novel gene trs1 encodes an essential protein for the transition from mitotic cell cycle to resting state in Schizosaccharomyces pombe" FEMS Microbiol.Lett.84. 1-6 (1991)

Kitamura、K. 和 C.Shimoda：“新基因 trs1 编码粟酒裂殖酵母从有丝分裂细胞周期到静息状态转变的必需蛋白质”FEMS Microbiol.Lett.84。

下田 親: "分裂酵母における体細胞分裂から減数分裂ヘの制御" 実験医学. 10. 171-176 (1992)

Chika Shimoda：“裂变酵母中体细胞分裂到减数分裂的控制”实验医学。10。171-176（1992）

T.Aono: "Mating pheromone-induced expression of the matl-Pm gene of Schizosaccharomomyces pombe:Identification of signalling components and characterisation of upstream controlling elements" YEAST. (in press).

T.Aono：“交配信息素诱导的粟酒裂殖酵母 matl-Pm 基因的表达：信号成分的鉴定和上游控制元件的表征”酵母。