Petersen J, Weilguny D, Egel R, Nielsen O
Department of Genetics, University of Copenhagen, Denmark.
Mol Cell Biol. 1995 Jul;15(7):3697-707. doi: 10.1128/MCB.15.7.3697.
In Schizosaccharomyces pombe, the fus1 mutation blocks conjugation at a point after cell contact and agglutination. The cell walls separating the mating partners are not degraded, which prevents cytoplasmic fusion. In order to investigate the molecular mechanism of conjugation, we cloned the fus1 gene and found that it is capable of encoding a 1,372-amino-acid protein with no significant similarities to other known proteins. Expression of the fus1 gene is regulated by the developmental state of the cells. Transcription is induced by nitrogen starvation and requires a pheromone signal in both P and M cell types. Consequently, mutants defective in the pheromone response pathway fail to induce fus1 expression. The ste11 gene, which encodes a transcription factor controlling expression of many genes involved in sexual differentiation, is also required for transcription of fus1. Furthermore, deletion of two potential Ste11 recognition sites in the fus1 promoter region abolished transcription, and expression could be restored when we inserted a different Ste11 site from the mat1-P promoter. Since this element was inverted relative to the fus1 element, we conclude that activation of transcription by Ste11 is independent of orientation. Although the fus1 mutant has a phenotype very similar to that of Saccharomyces cerevisiae fus1 mutants, the two proteins appear to have different roles in the process of cell fusion. Budding yeast Fus1 is a typical membrane protein and contains an SH3 domain. Fission yeast Fus1 has no features of a membrane protein, yet it appears to localize to the projection tip. A characteristic proline-rich potential SH3 binding site may mediate interaction with other proteins.
在粟酒裂殖酵母中,fus1突变在细胞接触和凝集后的某个点阻断接合过程。分隔交配伴侣的细胞壁未被降解,这阻止了细胞质融合。为了研究接合的分子机制,我们克隆了fus1基因,发现它能够编码一种1372个氨基酸的蛋白质,与其他已知蛋白质没有明显的相似性。fus1基因的表达受细胞发育状态的调节。转录由氮饥饿诱导,并且在P型和M型细胞中都需要信息素信号。因此,信息素反应途径有缺陷的突变体无法诱导fus1表达。ste11基因也参与fus1的转录,该基因编码一种控制许多参与性别分化基因表达的转录因子。此外,fus1启动子区域中两个潜在的Ste11识别位点的缺失消除了转录,当我们从matl-P启动子插入一个不同的Ste11位点时,可以恢复表达。由于该元件相对于fus1元件是反向的,我们得出结论,Ste11对转录的激活与方向无关。虽然fus1突变体的表型与酿酒酵母fus1突变体非常相似,但这两种蛋白质在细胞融合过程中似乎具有不同的作用。芽殖酵母Fus1是一种典型的膜蛋白,含有一个SH3结构域。裂殖酵母Fus1没有膜蛋白的特征,但它似乎定位于突起顶端。一个富含脯氨酸的潜在SH3结合位点可能介导与其他蛋白质的相互作用。