Alonso-Nuñez Maria Luisa, An Hanbing, Martín-Cuadrado Ana Belén, Mehta Sapna, Petit Claudia, Sipiczki Matthias, del Rey Francisco, Gould Katheleen L, de Aldana Carlos R Vázquez
Departamento de Microbiología y Genética, Instituto de Microbiología Bioquímica, CSIC/Universidad de Salamanca, Campus Miguel de Unamuno, Spain.
Mol Biol Cell. 2005 Apr;16(4):2003-17. doi: 10.1091/mbc.e04-06-0442. Epub 2005 Feb 2.
Schizosaccharomyces pombe cells divide by medial fission through contraction of an actomyosin ring and deposition of a multilayered division septum that must be cleaved to release the two daughter cells. Here we describe the identification of seven genes (adg1(+), adg2(+), adg3(+), cfh4(+), agn1(+), eng1(+), and mid2(+)) whose expression is induced by the transcription factor Ace2p. The expression of all of these genes varied during the cell cycle, maximum transcription being observed during septation. At least three of these proteins (Eng1p, Agn1p, and Cfh4p) localize to a ring-like structure that surrounds the septum region during cell separation. Deletion of the previously uncharacterized genes was not lethal to the cells, but produced defects or delays in cell separation to different extents. Electron microscopic observation of mutant cells indicated that the most severe defect is found in eng1Delta agn1Delta cells, lacking the Eng1p endo-beta-1,3-glucanase and the Agn1p endo-alpha-glucanase. The phenotype of this mutant closely resembled that of ace2Delta mutants, forming branched chains of cells. This suggests that these two proteins are the main activities required for cell separation to be completed.
粟酒裂殖酵母细胞通过肌动球蛋白环的收缩和多层分裂隔膜的沉积进行中间分裂,该隔膜必须被裂解才能释放出两个子细胞。在此,我们描述了七个基因(adg1(+)、adg2(+)、adg3(+)、cfh4(+)、agn1(+)、eng1(+)和mid2(+))的鉴定,其表达由转录因子Ace2p诱导。所有这些基因的表达在细胞周期中均有变化,在隔膜形成期间观察到最大转录。这些蛋白质中至少有三种(Eng1p、Agn1p和Cfh4p)定位于细胞分离期间围绕隔膜区域的环状结构。缺失先前未鉴定的基因对细胞并非致命,但在不同程度上导致细胞分离缺陷或延迟。对突变细胞的电子显微镜观察表明,在eng1Delta agn1Delta细胞中发现了最严重的缺陷,这些细胞缺乏Eng1p内切-β-1,3-葡聚糖酶和Agn1p内切-α-葡聚糖酶。该突变体的表型与ace2Delta突变体的表型非常相似,形成细胞分支链。这表明这两种蛋白质是完成细胞分离所需的主要活性物质。